Novel chimeric nucleic acids, encoding chimeric Borrelia proteins comprising OspC or an antigenic fragment thereof and OspA or an antigenic fragment thereof, are disclosed. Chimeric proteins encoded by the nucleic acid sequences are also disclosed. The chimeric proteins are useful as vaccine immunogens against Lyme borreliosis, as well as for immunodiagnostic reagents.

Current U.S. Class:	536/23.7 ; 424/243.1; 424/9.1; 424/9.2; 530/300; 530/350; 536/23.1
Current International Class:	C07H 21/04 (20060101); A61K 39/02 (20060101); C07H 21/02 (20060101)
Field of Search:	536/23.1,23.7 530/300,350 424/9.1,9.2,234.1

Hamilton, Brook, Smith & Reynolds, P.C.

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Government Interests

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GOVERNMENT SUPPORT

The invention was supported, in whole or in part, by Grant 2R01AI37256-05A1 from the National Institute of Allergy and Infectious Diseases. The Government has certain rights in the invention.

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Parent Case Text

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RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No. 11/196,475, filed Aug. 3, 2005, now U.S. Pat. No. 7,179,448, which is a divisional of U.S. application Ser. No. 10/369,100, filed Feb. 18, 2003, now U.S. Pat. No. 7,008,625, which is a continuation of International Application No. PCT/US01/24736, filed Aug. 7, 2001, published in English, which claims the benefit of U.S. Provisional Application No. 60/226,484, filed Aug. 18, 2000, said International Application No. PCT/US01/24736 also being a continuation-in-part of U.S. application Ser. No. 09/666,017, filed Sep. 19, 2000 (abandoned), which is itself a continuation-in-part of U.S. application Ser. No. 08/235,836 (now U.S. Pat. No. 6,248,562), filed Apr. 29, 1994, which is itself a continuation-in-part of U.S. application Ser. No. 08/148,191, filed Nov. 1, 1993 (abandoned). The entire teachings of the above application(s) are incorporated herein by reference.

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Claims

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What is claimed is:

1. A nucleic acid encoding a protein comprising at least a first and a second polypeptide, wherein the first polypeptide comprises OspC from Borrelia burgdorferi sensu lato and wherein the second polypeptide comprises OspA from Borrelia burgdorferi sensu lato, such that the first polypeptide is linked to the N-terminus of the second polypeptide.

2. A nucleic acid encoding a protein comprising at least a first and a second polypeptide, wherein the first polypeptide comprises OspC from Borrelia burgdorferi sensu lato from amino acid residue 19 to amino acid residue 211, wherein amino acid numbering is according to the OspC polypeptide numbering of SEQ ID NO: 30, and wherein the second polypeptide comprises OspA from Borrelia burgdorferi sensu lato selected from the group consisting of: (a) a Borrelia burgdorferi sensu lato OspA polypeptide from amino acid residue 18 to amino acid residue 273, (b) a Borrelia burgdorferi sensu lato OspA polypeptide from amino acid residue 132 to amino acid residue 216, (c) a Borrelia burgdorferi sensu lato polypeptide comprising at least two separate OspA polypeptide fragments, wherein the first OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 18 to amino acid residue 216 from a first strain of Borrelia burgdorferi sensu lato, and wherein the second OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 217 to amino acid residue 273 from a second strain of Borrelia burgdorferi sensu lato, (d) a Borrelia burgdorferi sensu lato polypeptide comprising at least two separate OspA polypeptide fragments, wherein the first OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 132 to amino acid residue 216 from a first strain of borrelia burgdorferi sensu lato, and wherein the second OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 217 to amino acid residue 273 from a second strain of Borrelia burgdorferi sensu lato, and (e) a Borrelia burgdorferi sensu lato polypeptide comprising at least four separate OspA polypeptide fragments, wherein the first OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 30 to amino acid residue 150, wherein the second OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 151 to amino acid residue 179, wherein the third OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 181 to amino acid residue 216 and, wherein the fourth OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 216 to amino acid residue 273, wherein at least two of the OspA polypeptide fragments are from different strains of Borrelia burgdorferi sensu lato, wherein amino acid numbering of OspA polypeptide is according to the OspA polypeptide numbering of SEQ ID NO: 7.

3. A nucleic acid encoding a protein comprising at least a first and a second polypeptide, wherein the first polypeptide comprises OspC from Borrelia burgdorferi sensu lato from amino acid residue 19 to amino acid residue 204, wherein amino acid numbering is according to the OspC polypeptide numbering of SEQ ID NO: 30, and wherein the second polypeptide comprises OspA from Borrelia burgdorferi sensu lato selected from the group consisting of: (a) a Borrelia burgdorferi sensu lato OspA polypeptide from amino acid residue 18 to amino acid residue 273, (b) a Borrelia burgdorferi sensu lato OspA polypeptide from amino acid residue 132 to amino acid residue 216, (c) a Borrelia burgdorferi sensu lato polypeptide comprising at least two separate OspA polypeptide fragments, wherein the first OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 18 to amino acid residue 216 from a first strain of Borrelia burgdorferi sensu lato, and wherein the second OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 217 to amino acid residue 273 from a second strain of Borrelia Borrelia sensu lato, (d) a Borrelia burgdorferi sensu lato polypeptide comprising at least two separate OspA polypeptide fragments, wherein the first OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 132 to amino acid residue 216 from a first strain of Borrelia burgdorferi sensu lato, and wherein the second OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 217 to amino acid residue 273 from a second strain of Borrelia burgdorferi sensu lato, and (e) a Borrelia burgdorferi sensu lato polypeptide comprising at least four separate OspA polypeptide fragments, wherein the first OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 30 to amino acid residue 150, wherein the second OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 151 to amino acid residue 179, wherein the third OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 181 to amino acid residue 216 and, wherein the fourth OspA polypeptide fragment comprises an OspA polypeptide from amino acid residue 216 to amino acid residue 273, wherein at least two of the OspA polypeptide fragments are from different strains of Borrelia burgdorferi sensu lato, wherein amino acid numbering of OspA polypeptide is according to the OspA polypeptide numbering of SEQ ID NO: 7.

4. A nucleic acid of any one of claims 1, 2, or 3, wherein the first polypeptide is from a first strain of Borrelia burgdorferi sensu lato and the second polypeptide is from a second strain of Borrelia burgdorferi sensu lato.

5. A nucleic acid of any one of claims 1, 2, or 3, wherein the first polypeptide is from a first genospecies of Borrelia burgdorferi sensu lato and the second polypeptide is from a second genospecies of Borrelia burgdorferi sensu lato.

6. A nucleic acid selected from the group consisting of: SEQ ID NOs: 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177 and 179.

7. An expression vector comprising an isolated DNA having a nucleotide sequence selected from the group consisting of: SEQ ID NOs: 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177 and 179.

8. An expression vector comprising an isolated DNA encoding a polypeptide having an amino acid sequence selected from the group consisting of: SEQ ID NOs: 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178 and 180.

9. A host cell comprising a recombinant nucleic acid having a nucleotide sequence selected from the group consisting of: SEQ ID NOs: 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177 and 179.

10. A host cell comprising a recombinant nucleic acid encoding a protein having an amino acid sequence selected from the group consisting of: SEQ ID NOs: 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178 and 180.

11. A physiological composition to vaccinate against and treat Borrelia infection in animals or humans, the composition comprising; a) a nucleic acid that encodes a chimeric protein comprising at least a first and a second polypeptide, wherein the first polypeptide comprises OspC from Borrelia burgdorferi sensu lato and wherein the second polypeptide comprises OspA from Borrelia burgdorferi sensu lato, such that the first polypeptide is linked to the N-terminus of the second polypeptide; and b) a physiologically-acceptable carrier or vehicle.

12. The composition of claim 11, wherein the nucleic acid is selected from the group consisting of: SEQ ID NOs: 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177 and 179.

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Description

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BACKGROUND OF THE INVENTION

Lyme disease (Lyme borreliosis) is the most common tick-borne infectious disease in North America and Europe, and has been found in Russia, Japan, China and Australia. Lyme disease begins at the site of a tick bite, producing a primary infection with spread of the organism to secondary sites occurring during the course of infection. The causative bacterial agent of this disease is the spirochete Borrelia burgdorferi, which was first isolated and cultivated in 1982 (Burgdorferi, W. A. et al., Science 216: 1317-1319 (1982); Steere, A. R. et al., N. Engl. J. Med. 308: 733-740 (1983)). With that discovery, a wide array of clinical syndromes, described in both the European and American literature since the early 20th century, could be attributed to infection by B. burgdorferi (Afzelius, A., Acta Derm. Venereol. 2: 120-125 (1921); Bannwarth, A., Arch. Psychiatr. Nervenkrankh. 117: 161-185 (1944); Garin, C. and A. Bujadouz, J. Med. Lyon 71: 765-767 (1922); Herxheimer, K. and K. Hartmann, Arch. Dermatol. Syphilol. 61: 57-76, 255-300 (1902)).

Three pathogenic genospecies of Borrelia, B. burgdorferi sensu stricto (B. burgdorferi or B.b.s.s.), B. afzelii and B. garinii have been described (Baranton, G., et al., Int. J. Syst. Bacteriol. 42:378-383 (1992)). These are members of a species complex, B. burgdorferi sensu lato, which consists of at least 10 different genospecies (Piken, R. N., et al., J. Invest. Dermatol., 110:211-214 (1998); Postic, D., et al., Int. J. Syst. Bacteriol. 44:743-752 (1994); Valsangiacomo, C. T., et al., Int. J. Syst. Bacteriol. 47:1-10 (1997)). The three genospecies, B. burgdorferi sensu stricto, B. afzelii and B. garinii, all are thought to be pathogenic and all are found in Europe. However, in North America, B. burgdorferi sensu stricto is the only identified pathogenic genospecies. Each of these three genospecies is associated with distinct clinical manifestations (Van Dam, A. P. et al., Clin. Infect. Dis. 17:708-717 (1993)). This implies that differences in genospecies may play an important role in the wide array of clinical manifestations observed in Lyme Disease.

OspA is a basic lipoprotein of approximately 31 kd, which is encoded on a large linear plasmid along with OspB, a basic lipoprotein of approximately 34 kd (Szczepanski, A., and J. L. Benach, Microbiol. Rev. 55:21 (1991)). Analysis of isolates of B. burgdorferi obtained from North America and Europe has demonstrated that OspA has antigenic variability, and that several distinct groups can be serologically and genotypically defined (Wilske, B., et al., World J. Microbiol. 7: 130 (1991)). Other Borrelia proteins demonstrate similar antigenic variability. Surprisingly, the immune response to these outer surface proteins tends to occur late in the disease, if at all (Craft, J. E. et al., J. Clin Invest. 78: 934-939 (1986); Dattwyler, R. J. and B. J. Luft, Rheum. Clin. North Am. 15: 727-734 (1989)). Furthermore, patients acutely and chronically infected with B. burgdorferi respond variably to the different antigens, including OspA, OspB, OspC, OspD, p39, p41 and p93.

As an infected tick begins to feed on a mammal, the synthesis of another outer surface protein, outer surface protein C or OspC, is induced (Schwan, T. G., et al., Proc. Natl. Acad. Sci. 2:2909-2913 (1995)). Thus, in early infection, OspC is the major outer membrane protein expressed by the spirochete (Fung, B. P., et al., Infect. Immun. 62:3213-3221 (1994); Padula, S. J., et al., J. Clin. Microbiol., 32:1733-1738 (1994)). Even through OspC has been demonstrated to have limited surface exposure (Cox, D. L., et al., Proc. Natl. Acad. Sci., 93:7973-7978 (1996); Mathiesen, M. M., et al., Infect. Immun. 66:4073-4079 (1998)), OspC is a potent immunogen. Immunization with OspC is protective against tick-transmitted Borrelia infection (Gilmore Jr., R. D., Infect. Immun. 64:2234-2239 (1999)). However, because OspC is highly variable in its sequence, the protection is limited to the Borrelia burgdorferi strain expressing the same allele of OspC. Challenge with heterologous isolates, expressing other ospC alleles results in infection (Probert, W. S., et al., J. Infect. D., 175:400-405 (1997)). OspC is a very diverse genetic locus (Jauris-Heipke, S., et al., Med. Microbiol. Immunol. 182:37-50 (1993)) as evidenced by the fact that Livey et al. found thirty-four alleles of OspC in seventy-six B. burgdorferi sensu lato isolates (Livey, I., et al., Mol. Microbiol. 18:257-269 (1995)).

Currently, Lyme Disease is treated with a range of antibiotics, e.g., tetracyclines, penicillin and cephalosporins. However, such treatment is not always successful in clearing the infection. Treatment is often delayed due to improper diagnosis with the deleterious effect that the infection proceeds to a chronic condition, where treatment with antibiotics is often not useful. One of the factors contributing to delayed treatment is the lack of effective diagnostic tools.

Vaccines against Lyme borreliosis have been attempted. Mice immunized with a recombinant form of OspA are protected from challenge with the same strain of B. burgdorferi from which the protein was obtained (Fikrig, E., et al., Science 250: 553-556 (1990)). Furthermore, passively transferred anti-OspA monoclonal antibodies (MAbs) have been shown to be protective in mice, and vaccination with a recombinant protein induced protective immunity against subsequent infection with the homologous strain of B. burgdorferi (Simon, M. M., et al., J. Infect. Dis. 164: 123 (1991)). In addition, there have been two independent trials of first generation vaccines for the prevention of Lyme disease that have studied the efficacy and safety of a vaccine based on recombinant outer surface protein A (OspA) (Sigal, L. H. et al., N. Engl. J. Med 339:216-222, 1998; Steere, A. C. et al., N. Engl. J. Med. 339:209-215, (1998)). However, a vaccine that consists of recombinant OspA may require frequent booster immunizations. An additional concern of OspA-based vaccines is the recent identification of a putative autoreactive OspA domain with a high degree of similarity to a region of human leukocyte function-associated antigen-1 (hLFA-1) (Gross, D. M. et al., Science, 281: 703-706 (1998)).

It has been noted that immunization with a single protein from a particular strain of Borrelia often does not confer resistance to that strain in all individuals (Fikrig, E. et al., J. Immunol. 7: 2256-1160 (1992)). There is considerable variation displayed in OspA, OspB and OspC, as well as p93, including the regions conferring antigenicity. Therefore, the degree and frequency of protection from vaccination with a protein from a single strain depend upon the response of the immune system to the particular variation, as well as the frequency of genetic variation in B. burgdorferi. In the case of vaccines directed against OspA, the vaccine is typically only effective against strains of Borrelia that express OspA that is homologous to OspA from which the vaccine was derived.

Another limitation of current OspA Lyme Disease vaccines is that they are directed against an antigen that is expressed predominantly in the tick vector. Indeed, recent reports have indicated that Borrelia burgdorferi in infected ticks alter their surface expression by increasing expression of OspC during ingestion of a blood meal (Schwan, T. G. et al., Proc. Natl. Acad. Sci. USA, 92: 2909-2913 (1995)). Thus, it seems that natural infection with B. burgdorferi does not elicit an antibody response to OspA, as it does against OspC.

Given the heterogeneity of antigenic determinants present in Borrelia proteins, a need exists for a vaccine and diagnostic tool which can provide immunogenicity to various strains and/or genospecies of Borrelia burgdorferi, as well as to more epitopes within a strain or genospecies. There is also a need for vaccines and diagnostic tools which detect antibody responses against immunoprotective targets that are expressed at different stages of the life cycle of Borrelia burgdorferi. This would allow for diagnosis and/or vaccination against all, or most forms, of Borrelia that cause systemic disease.

SUMMARY OF THE INVENTION

The current invention pertains to chimeric Borrelia proteins which include two or more antigenic Borrelia polypeptides which do not occur naturally (in nature) in the same protein in Borrelia, as well as the nucleic acids encoding such chimeric proteins. The proteins from which the antigenic polypeptides are derived can be from the same strain or genospecies of Borrelia, from different strains or genospecies, or from combinations of proteins from the same and from different strains or genospecies. Particular chimeric proteins, and the nucleotide sequences encoding them, are set forth in FIGS. 30-37 and 55-72.

The chimeric proteins of the current invention provide antigenic polypeptides of a variety of Borrelia strains and/or proteins within a single protein. Such proteins are particularly useful in immunodiagnostic assays to detect the presence of antibodies to native Borrelia in potentially infected individuals as well as to measure T-cell reactivity, and can therefore be used as immunodiagnostic reagents. These chimeric proteins are also useful in the generation of immune responses (such as antibody production) against proteins expressed by Borrelia burgdorferi. The chimeric proteins of the current invention are additionally useful as vaccine immunogens against Borrelia infection.

In one embodiment of the present invention, the chimeric proteins are made up of polypeptide fragments from Lyme Disease-causing strains of Borrelia. In another embodiment, the polypeptide fragments that make up the chimeric protein are from outer surface protein A (OspA) and outer surface protein C (OspC), which have the general structure of OspC linked via a peptide bond to the N-terminus of OspA. The present invention encompasses both lipidated and unlipidated chimeric proteins. In one embodiment, the OspA and OspC portions of the chimeric protein possess a lipidation signal. In other embodiments, either the OspA polypeptide portion, the OspC polypeptide portion, or both, do not include a lipidation signal.

The OspA portion of the chimeric polypeptide can itself comprise OspA portions from two or more strains of Lyme Disease-causing Borrelia as described herein and provided, for example, in FIGS. 23-29 and 43-46. In another embodiment, the OspA polypeptide comprises OspA portions from two or more genospecies of Lyme Disease-causing Borrelia, for example, wherein the genospecies are defined as Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii. In this manner, the OspC and OspA polypeptide fragments that make up the chimeric protein can be from the same strain or genospecies of Borrelia, from different strains or genospecies of Borrelia, or from combinations of proteins from the same and from different strains or genospecies of Borrelia.

The present invention is also drawn to nucleic acids which encode a Borrelia chimeric protein. In a particular embodiment, the composition comprises a nucleic acid that encodes a chimeric protein of at least two polypeptides, wherein the first polypeptide comprises Borrelia burgdorferi OspC, and the second polypeptide comprises Borrelia burgdorferi OspA, such that OspC is upstream of OspA. The OspC and OspA nucleic acid fragments that make up the chimeric protein can be from the same strain or genospecies of Borrelia, from different strains or genospecies of Borrelia, or from combinations of proteins that are from the same and/or different strains or genospecies of Borrelia.

The present invention is also drawn to expression vectors which comprise an isolated DNA encoding a Borrelia chimeric protein. In one embodiment, the composition includes an expression vector comprising an isolated DNA which encodes an OspC/OspA chimeric protein as described herein. The present invention also encompasses host cells which comprise a recombinant nucleic acid encoding an OspC/OspA chimeric protein as described herein.

The present invention is also drawn to methods of making the Borrelia chimeric polypeptides described herein. In one embodiment, the method of making a chimeric Borrelia protein comprises selecting a polynucleotide sequence encoding OspC, or an antigenic portion thereof, selecting a polynucleotide sequence encoding OspA, or an antigenic portion thereof, and ligating these polynucleotide sequences together.

The present invention is also drawn to methods of delivering the Borrelia chimeric polypeptides described herein. In one embodiment, the method comprises administering the chimeric protein in a physiologically-acceptable carrier to an individual. As a result of the administration of the chimeric protein, the individual develops at least some immune response to the chimeric protein, e.g., the individual generates a humoral immune response, wherein antibodies are produced by the individual that recognize at least a portion of said chimeric polypeptide.

The present invention is also drawn to methods of delivering nucleic acids which encode the chimeric polypeptides described herein. In one embodiment, the method comprises administering the nucleic acid in a physiologically-acceptable carrier to an individual. As a result of the administration of the nucleic acid, the individual expresses the chimeric protein at least transiently and develops at least some immune response to the chimeric protein encoded by the nucleic acid, e.g., the individual generates a humoral immune response, wherein antibodies that recognize at least a portion of the chimeric polypeptide produced from the nucleic acid, are produced by the individual.

The invention also encompasses methods of using the chimeric proteins described herein in a diagnostic assay. As described herein, the method can be used to detect the presence of OspA- and/or OspC-specific antibodies in a sample, e.g., a host sample of interest. The method comprises contacting a sample, e.g., a host sample of interest, with the chimeric protein, under conditions, wherein antibodies, if present in the host sample, bind to the chimeric protein thereby forming antigen-antibody complexes. The antigen-antibody complexes are then detected. In this manner, the chimeric proteins of the present invention can be used to detect an immune response to Lyme Disease causing Borrelia.

The present invention is also drawn to diagnostic kits which comprise the chimeric polypeptides described herein. In one embodiment, the kit comprises a Borrelia burgdorferi OspC/OspA chimeric protein. The kit also includes reagents for detecting antibody-antigen complexes that are formed between the OspC/OspA chimeric protein and antibodies that are present in a sample, e.g., a user-supplied host sample.

The present invention is also drawn to pharmaceutical compositions which can be used to vaccinate and/or treat Borrelia infection in an animal or human. The pharmaceutical composition can be administered together with a physiologically-acceptable carrier and/or with suitable excipients and/or adjuvants.

The present invention is also drawn to methods of immunizing an animal or human against Lyme disease. In a particular embodiment, the method comprises administering a Borrelia chimeric OspC/OspA protein. The chimeric protein can be administered together with a physiologically-acceptable carrier, a suitable excipient and/or a suitable adjuvant, to an animal or human such that the animal or human develops an immune response to at least one of the OspC and/or OspA polypeptides of the composition.

By incorporating polypeptide fragments from multiple Borrelia burgdorferi proteins, the present invention provides a composition that has great utility for vaccines and diagnostic kits. As a result of the present invention, there exist diagnostic tools and vaccines that comprise both OspA and OspC antigens from various Borrelia burgdorferi strains and/or genospecies in a single protein. Since OspA is primarily expressed in the tick vector, and OspC is upregulated in response to the feeding of an infected tick on a mammal, this allows for a diagnostic tool or vaccine that can recognize antigens that are expressed at different stages of the life cycle of Borrelia burgdorferi. Thus, the chimeric proteins of the present invention can act at the level of the tick as well as the level of the host, in preventing infection and/or disease caused by Borrelia. Moreover, by incorporating unique polypeptide fragments from pathogenic families of Borrelia, such as Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii, an improved diagnostic tool or vaccine is produced which can detect clinically important exposure to a wider variety of pathogenic Borrelia, while overlooking the remainder of non-pathogenic families of Borrelia. Furthermore, OspC polypeptides can be selected from strains of Borrelia that are associated with disseminated disease, as described in WO 00/78966, the teachings of which are incorporated herein in their entirety.

The present invention also provides a combination of Borrelia antigens in a single polypeptide that, when used as a vaccine, are expected to prevent Lyme disease from becoming systemic. The chimeric proteins of the present invention can be effective in preventing Lyme disease, as well as having a therapeutic effect on established infection, for example after the tick bite is noticed by the patient.

The present invention is drawn to both lipidated and unlipidated chimeric proteins. Unlipidated chimeric proteins, such as the OspC/OspA chimeric proteins described herein, have certain advantages over their lipidated counterparts. These advantages include simpler production methods, improved yields of protein and simpler purification methods. While the lack of a lipidation signal provides several advantages, it had been thought that the lipidation signal was required for immunogenicity. However, as described herein, the non-lipidated OspC/OspA chimeric proteins of the present invention elicit an immune response that is at least as broadly reactive as that of lipidated OspA and lipidated OspC control proteins. Moreover, the unlipidated OspC/OspA chimeric proteins of the present invention unexpectedly elicit an immune response to more than one genospecies and/or strain of Lyme disease-causing Borrelia, including genospecies and/or strains that were not used to generate the particular chimeric OspC/OspA immunogen.

For a better understanding of the present invention together with other and further objects, reference is made to the following description, taken together with the accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 summarizes peptides and antigenic domains localized by proteolytic and chemical fragmentation of OspA.

FIG. 2 is a comparison of the antigenic domains depicted in FIG. 1, for OspA in nine strains of B. burgdorferi. In Domain 1, A-B31 is SEQ ID NO: 187, A-TRo is SEQ ID NO: 187; A-K48 is SEQ ID NO: 188; A-DK29 is SEQ ID NO: 188; A-P/Gau is SEQ ID NO: 187; A-PKo is SEQ ID NO: 187; A-IP3 is SEQ ID NO: 189; A-IP90 is SEQ ID NO: 194; and A-25015 is SEQ ID NO: 187. In Domain 2, A-B31 is SEQ ID NO: 191, A-TRo is SEQ ID NO: 192; A-K48 is SEQ ID NO: 193; A-DK29 is SEQ ID NO: 193; A-P/Gau is SEQ ID NO: 194; A-PKo is SEQ ID NO: 194; A-IP3 is SEQ ID NO: 195; A-IP90 is SEQ ID NO: 192; and A-25015 is SEQ ID NO: 191. In Domain 3, A-B31 is SEQ ID NO: 196, A-TRo is SEQ ID NO: 197; A-K48 is SEQ ID NO: 198; A-DK29 is SEQ ID NO: 199; A-P/Gau is SEQ ID NO: 200; A-PKo is SEQ ID NO: 200; A-IP3 is SEQ ID NO: 200; A-IP90 is SEQ ID NO: 201; and A-25015 is SEQ ID NO: 202. In Domain 4, A-B31 is SEQ ID NO: 203, A-TRo is SEQ ID NO: 294; A-K48 is SEQ ID NO: 205; A-DK29 is SEQ ID NO: 205; A-P/Gau is SEQ ID NO: 206; A-PKo is SEQ ID NO: 206; A-IP3 is SEQ ID NO: 206; A-IP90 is SEQ ID NO: 205; and A-25015 is SEQ ID NO: 206.

FIG. 3 is a graph depicting a plot of weighted polymorphism versus amino acid position among 14 OspA variants. The marked peaks are: a) amino acids 132-145; b) amino acids 163-177; c) amino acids 208-221. The lower line at polymorphism value 1.395 demarcates statistically significant excesses of polymorphism at p=0.05. The upper line at polymorphism value 1.520 is the same, except that the first 29 amino acids at the monomorphic N-terminus have been removed from the original analysis.

FIG. 4 depicts the amino acid alignment of residues 200 through 220 for OspAs from strains B31 (SEQ ID NO: 207) and K48 (SEQ ID NO: 208) as well as for the site-directed mutants 613 (SEQ ID NO: 209), 625 (SEQ ID NO: 210), 640 (SEQ ID NO: 211), 613/625 (SEQ ID NO: 212), and 613/640 (SEQ ID NO: 213). The arrow indicates Trp216. Amino acid changes are underlined.

FIG. 5 is a helical wheel projection of residues 204-217 of B31 OspA. Capital letters indicate hydrophobic residues; lower case letters indicate hydrophilic residues; + and - indicate positively and negatively charged residues, respectively. The dashed line indicates division of the alpha-helix into a hydrophobic arc (above the line) and a polar arc (below the line). Adapted from France et al. (Biochem. Biophys. Acta 1120: 59 (1992)).

FIG. 6 depicts a phylogenic tree for strains of Borrelia described in Table I. The strains are as follows: 1=B31; 2=PKa1; 3=ZS7; 4=N40; 5=25015; 6=K48; 7=DK29; 8=PHei; 9=Ip90; 10=PTrob; 11=ACAI; 12=PGau; 13=Ip3; 14=PBo; 15=PKo.

FIGS. 7A and 7B depict the nucleic acid sequence of OspA-B31 (SEQ ID NO. 6), and the encoded protein sequence (SEQ ID NO. 7).

FIGS. 8A, 8B and 8C depict the nucleic acid sequence of OspA-K48 (SEQ ID NO. 8), and the encoded protein sequence (SEQ ID NO. 9).

FIGS. 9A, 9B and 9C depict the nucleic acid sequence of OspA-PGau (SEQ ID NO. 10), and the encoded protein sequence (SEQ ID NO. 11).

FIGS. 10A and 10B depict the nucleic acid sequence of a portion of an OspA gene (SEQ ID NO. 185) and its encoded protein sequence (SEQ ID NO. 186).

FIGS. 11A, 11B and 11C depict the nucleic acid sequence of OspB-B31 (SEQ ID NO. 21), and the encoded protein sequence (SEQ ID NO. 22).

FIGS. 12A and 12B depict the nucleic acid sequence of OspC-B31 (SEQ ID NO. 29), and the encoded protein sequence (SEQ ID NO. 30).

FIGS. 13A and 13B depict the nucleic acid sequence of OspC-K48 (SEQ ID NO. 31), and the encoded protein sequence (SEQ ID NO. 32).

FIGS. 14A and 14B depict the nucleic acid sequence of OspC-PKo (SEQ ID NO. 33), and the encoded protein sequence (SEQ ID NO. 34).

FIGS. 15A and 15B depict the nucleic acid sequence of OspC-PTrob (SEQ ID NO. 35) and the encoded protein sequence (SEQ ID NO. 36).

FIGS. 16A, 16B, 16C, 16D and 16E depict the nucleic acid sequence of p93-B31 (SEQ ID NO. 65) and the encoded protein sequence (SEQ ID NO. 66).

FIG. 17 depicts the nucleic acid sequence of p93-K48 (SEQ ID NO. 67).

FIG. 18 depicts the nucleic acid sequence of p93-PBo (SEQ ID NO. 69).

FIG. 19 depicts the nucleic acid sequence of p93-PTrob (SEQ ID NO. 71).

FIG. 20 depicts the nucleic acid sequence of p93-PGAU (SEQ ID NO. 73).

FIG. 21 depicts the nucleic acid sequence of p93-25015 (SEQ ID NO. 77).

FIG. 22 depicts the nucleic acid sequence of p93-PKo (SEQ ID NO. 75).

FIGS. 23A, 23B and 23C depict the nucleic acid sequence of the OspA-K48/OspA-PGAU chimer (SEQ ID NO. 85) and the encoded chimeric protein sequence (SEQ ID NO. 86).

FIGS. 24A, 24B and 24C depict the nucleic acid sequence of the OspA-B31/OspA-PGAU chimer (SEQ ID NO. 88) and the encoded chimeric protein sequence (SEQ ID NO. 89).

FIGS. 25A and 25B depict the nucleic acid sequence of the OspA-B31/OspA-K48 chimer (SEQ ID NO. 91) and the encoded chimeric protein sequence (SEQ ID NO. 92).

FIGS. 26A, 26B and 26C depict the nucleic acid sequence of the OspA-B31/OspA-25015 chimer (SEQ ID NO. 94) and the encoded chimeric protein sequence (SEQ ID NO. 95).

FIGS. 27A, 27B and 27C depict the nucleic acid sequence of the OspA-K48/OspA-B31/OspA-K48 chimer (SEQ ID NO. 97) and the encoded chimeric protein sequence (SEQ ID NO. 98).

FIGS. 28A, 28B and 28C depict the nucleic acid sequence of the OspA-B31/OspA-K48/OspA-B31/OspA-K48 chimer (SEQ ID NO. 100) and the encoded chimeric protein sequence (SEQ ID NO. 101).

FIGS. 29A, 29B and 29C depict the nucleic acid sequence of the OspA-B31/OspB-B31 chimer (SEQ ID NO. 103) and the encoded chimeric protein sequence (SEQ ID NO. 104).

FIGS. 30A, 30B, 30C and 30D depict the nucleic acid sequence of the OspA-B31/OspB-B31/OspC-B31 chimer (SEQ ID NO. 106) and the encoded chimeric protein sequence (SEQ ID NO. 107).

FIGS. 31A, 31B, 31C and 31D depict the nucleic acid sequence of the OspC-B31/OspA-B31/OspB-B31 chimer (SEQ ID NO. 109) and the encoded chimeric protein sequence (SEQ ID NO. 110).

FIGS. 32A, 32B, 32C, 32D and 32E depict the nucleic acid sequence of the OspA-B31/p93-B31 chimer (SEQ ID NO. 111) and the encoded chimeric protein sequence (SEQ ID NO. 112).

FIGS. 33A, 33B, 33C and 33D depict the nucleic acid sequence of the OspB-B31/p41-B31 (122-234) chimer (SEQ ID NO. 113) and the encoded chimeric protein sequence (SEQ ID NO. 114).

FIGS. 34A, 34B, 34C and 34D depict the nucleic acid sequence of the OspB-B31/p41-B31 (122-295) chimer (SEQ ID NO. 115) and the encoded chimeric protein sequence (SEQ ID NO. 116).

FIGS. 35A, 35B and 35C depict the nucleic acid sequence of the OspB-B31/p41-B31 (140-234) chimer (SEQ ID NO. 117) and the encoded chimeric protein sequence (SEQ ID NO. 118).

FIGS. 36A, 36B, 36C and 36D depict the nucleic acid sequence of the OspB-B31/p41-B31 (140-295) chimer (SEQ ID NO. 119) and the encoded chimeric protein sequence (SEQ ID NO. 120).

FIGS. 37A, 37B, 37C, 37D and 37E depict the nucleic acid sequence of the OspB-B31/p41-B31 (122-234)/OspC-B31 chimer (SEQ ID NO. 121) and the encoded chimeric protein sequence (SEQ ID NO. 122).

FIGS. 38A, 38B, 38C and 38D depict an alignment of the nucleic acid sequences for OspC-B31 (SEQ ID NO. 29), OspC-PKo (SEQ ID NO. 33), OspC-PTrob (SEQ ID NO. 35), and OspC-K48 (SEQ ID NO. 31). Nucleic acids which are identical to those in the lead nucleic acid sequence (here, OspC-B31) are represented by a period (.); differing nucleic acids are shown in lower case letters.

FIGS. 39A, 39B, 39C and 39D depict an alignment of the nucleic acid sequences for OspD-PBo (SEQ ID NO. 123), OspD-PGAU (SEQ ID NO. 124), OspD-DK29 (SEQ ID NO. 125), and OspD-K48 (SEQ ID NO. 126). Nucleic acids which are identical to those in the lead nucleic acid sequence (here, OspD-PBo) are represented by a period (.); differing nucleic acids are shown in lower case letters.

FIGS. 40A, 40B and 40C depict the nucleic acid sequence of p41-B31 (SEQ ID NO. 127) and then encoded protein sequence (SEQ ID NO. 128).

FIGS. 41A, 41B, 41C, 41D, 41E, 41F, 41G and 41H depict an alignment of the nucleic acid sequences for p41-B31 (SEQ ID NO. 127), p41-PKa1 (SEQ ID NO. 129), p41-PGAU (SEQ ID NO. 51), p41-PBo (SEQ ID NO. 130), p41-DK29 (SEQ ID NO. 53), and p41-PKo (SEQ ID NO. 131). Nucleic acids which are identical to those in the lead nucleic acid sequence (here, p41-B31) are represented by a period (.); differing nucleic acids are shown in lower case letters.

FIGS. 42A, 42B, 42C, 42D, 42E, 42F, 42G, 42H, 42I, 42J, 42K, 42L, 42M, 42N, 42O and 42P depict an alignment of the nucleic acid sequences for OspA-B31 (SEQ ID NO. 6), OspA-PKa1 (SEQ ID NO. 132), OspA-N40 (SEQ ID NO. 133), OspA-ZS7 (SEQ ID NO. 134), OspA-25015 (SEQ ID NO. 12), OspA-PTrob (SEQ ID NO. 135), OspA-K48 (SEQ ID NO. 8), OspA-Hei (SEQ ID NO. 136), OspA-DK29 (SEQ ID NO. 49), OspA-Ip90 (SEQ ID NO. 50), OspA-PBo (SEQ ID NO. 55), OspA-Ip3 (SEQ ID NO. 56), OspA-PKo (SEQ ID NO. 57), OspA-ACAI (SEQ ID NO. 58), and OspA-PGAU (SEQ ID NO. 10). Nucleic acids which are identical to those in the lead nucleic acid sequence (here, OspA-B31) are represented by a period (.); differing nucleic acids are shown in lower case letters.

FIGS. 43A and 43B depict the nucleic acid sequence of the OspA-Tro/OspA-Bo chimer (SEQ ID NO. 137) which encodes the chimeric protein sequence SEQ ID NO. 138.

FIGS. 44A and 44B depict the nucleic acid sequence of the OspA-PGAU/OspA-Bo chimer (SEQ ID NO. 139) which encodes the chimeric protein sequence SEQ ID. NO. 140.

FIGS. 45A and 45B depict the nucleic acid sequence of the OspA-B31/OspA-PGAU/OspA-B31/OspA-K48 chimer (SEQ ID NO. 143) which encodes the chimeric protein sequence SEQ ID NO. 144.

FIGS. 46A and 46B depict the nucleic acid sequence of the OspA-PGAU/OspA-B31/OspA-K48 chimer (SEQ ID NO. 141) which encodes the chimeric protein sequence SEQ ID NO. 142.

FIG. 47 is a bar graph showing the reactivity (as measured by ELISA) of sera from mice immunized with the indicated Borrelia protein (OspA or OspC) or recombinant chimeric protein (OspC2-OspA) (X-axis) against OspA B31 or OspC B31 antigens (legend).

FIG. 48 is a bar graph showing the reactivity (as measured by ELISA) of sera from mice immunized with the indicated Borrelia protein (OspA or OspC) or recombinant chimeric protein (OspC2-OspA) (X-axis) against OspA B31 or OspC B31 antigens (legend). For the ELISA results to the B31 OspA antigen, a purified fragment of B31 OspA (amino acids 18-139) was added in excess to the sera so that the detected immune response was specific for the C-terminal region of OspA.

FIG. 49 is a bar graph showing the reactivity of sera from mice immunized with the indicated Borrelia chimeric protein (lipOspA/Bo, lipOspAB/P or OspC-OspAB/P) (X-axis) against the indicated OspA antigens (legend) from strains B31 (Borrelia burgdorferi sensu stricto), K48 (Borrelia garinii) and PGau (Borrelia afzelli).

FIG. 50 is a bar graph showing the reactivity of sera from mice immunized with the indicated Borrelia chimeric protein (lipOspAP/Bo, lipOspAB/P) or OspC-OspAB/P) (X-axis) against the indicated OspA (legend) from strains B31 (Borrelia burgdorferi sensu stricto), K48 (Borrelia garinii) and PGau (Borrelia afzelli). In all cases, a purified fragment of B31 OspA (amino acids 18-139) was added in excess to the sera so that the detected immune response is specific for the C-terminal region of OspA.

FIG. 51 is a bar graph showing the reactivity of sera from mice immunized with the indicated Borrelia chimeric protein (OspCB31-OspAB31, OspC2-OspAB31 or lip OspC-B31) (X-axis) against the indicated OspC antigen (legend) from the strain B31 (Borrelia burgdorferi sensu stricto).

FIG. 52 is a bar graph showing the reactivity of sera from mice immunized with the indicated Borrelia chimeric protein (OspCB31-OspAB31, OspC2-OspAB31 or Lip OspA K/T) (X-axis) against the indicated OspA antigens (legend) from strains B31 (Borrelia burgdorferi sensu stricto), K48 (Borrelia garinii) and PGau (Borrelia afzelli).

FIG. 53 is a bar graph showing the reactivity of sera from mice immunized with the indicated Borrelia chimeric protein (OspCB31-OspAB/P, OspCB31-OspABPBP or OspCB31-OspAB31) (X-axis) against the indicated OspA antigens (legend) from strains B31 (Borrelia burgdorferi sensu stricto), K48 (Borrelia garinii) and PGau (Borrelia afzelli).

FIG. 54 is a bar graph showing the reactivity of sera from mice immunized with the indicated Borrelia chimeric protein (OspCB31-OspAB/P, OspCB31-OspABPBP or OspCB31-OspAB31) (X-axis) against the indicated OspA (legend) from strains B31 (Borrelia burgdorferi sensu stricto), K48 (Borrelia garinii) and PGau (Borrelia afzelli). In all cases, a purified fragment of B31 OspA (amino acids 18-139) was added in excess to the sera so that the detected immune response is specific for the C-terminal region of OspA.

FIGS. 55A, 55B and 55C depict the nucleic acid sequence of the OspC-B31 (bp 55-633)/OspA-B31 (bp 52-822) chimer (SEQ ID NO. 145) and the encoded chimeric protein sequence (SEQ ID NO. 146).

FIGS. 56A, 56B and 56C depict the nucleic acid sequence of the OspC-B31 (bp 55-624)/OspA-B31 (bp 52-822) chimer (SEQ ID NO. 147) and the encoded chimeric protein sequence (SEQ ID NO. 148).

FIGS. 57A, 57B and 57C depict the nucleic acid sequence of the OspC-C2 (bp 55-612)/OspA-B31 (bp 52-822) chimer (SEQ ID NO. 149) and the encoded chimeric protein sequence (SEQ ID NO. 150).

FIGS. 58A, 58B and 58C depict the nucleic acid sequence of the OspC-B31 (bp 55-633)/OspA-B31 (bp 52-651)/OspA-K48 (bp 652-820) chimer (SEQ ID NO. 151) and the encoded chimeric protein sequence (SEQ ID NO. 152).

FIGS. 59A, 59B and 59C depict the nucleic acid sequence of the OspC-C2 (bp 55-612)/OspA-B31 (bp 52-651)/OspA-K48 (bp 652-820) chimer (SEQ ID NO. 153) and the encoded chimeric protein sequence (SEQ ID NO. 154).

FIGS. 60A, 60B and 60C depict the nucleic acid sequence of the OspC-B31 (bp 55-633)/OspA-B31 (bp 52-651)/OspA-PKo (bp 652-820) chimer (SEQ ID NO. 155) and the encoded chimeric protein sequence (SEQ ID NO. 156).

FIGS. 61A, 61B and 61C depict the nucleic acid sequence of the OspC-C2 (bp 55-612)/OspA-B31 (bp 52-651)/OspA-PKo (bp 652-820) chimer (SEQ ID NO. 157) and the encoded chimeric protein sequence (SEQ ID NO. 158).

FIGS. 62A, 62B and 62C depict the nucleic acid sequence of the OspC-B31 (bp 55-633)/OspA-K48 (bp 52-654)/OspA-Tro (bp 655-819) chimer (SEQ ID NO. 159) and the encoded chimeric protein sequence (SEQ ID NO. 160).

FIGS. 63A, 63B and 63C depict the nucleic acid sequence of the OspC-C2 (bp 55-612)/OspA-K48 (bp 52-654)/OspA-Tro (bp 655-819) chimer (SEQ ID NO. 161) and the encoded chimeric protein sequence (SEQ ID NO. 162).

FIGS. 64A, 64B and 64C depict the nucleic acid sequence of the OspC-C12 (bp 55-612)/OspA-B31 (bp 88-492)/OspA-PKo (bp 493-537)/OspA-B31 (bp 538-822) chimer (SEQ ID NO. 163) and the encoded chimeric protein sequence (SEQ ID NO. 164).

FIGS. 65A, 65B and 65C depict the nucleic acid sequence of the OspC-PKo (bp 55-639)/OspA-B31 (bp 88-492)/OspA-PKo (bp 493-537)/OspA-B31 (bp 538-651)/OspA-K48 (bp 652-825) chimer (SEQ ID NO. 165) and the encoded chimeric protein sequence (SEQ ID NO. 166).

FIGS. 66A, 66B and 66C depict the nucleic acid sequence of the OspC-Tro (bp 55-624)/OspA-B31 (bp 88-492)/OspA-PKo (bp 493-537)/OspA-B31 (bp 538-651)/OspA-PKo (bp 652-822) chimer (SEQ ID NO. 167) and the encoded chimeric protein sequence (SEQ ID NO. 168).

FIGS. 67A and 67B depict the nucleic acid sequence of the OspC-B31 (bp 55-633)/OspA-B31 (bp 394-820) chimer (SEQ ID NO. 169) and the encoded chimeric protein sequence (SEQ ID NO. 170).

FIGS. 68A and 68B depict the nucleic acid sequence of the OspC-B31 (bp 55-631)/OspA-B31 (bp 394-651)/OspA-K48 (bp 652-820) chimer (SEQ ID NO. 171) and the encoded chimeric protein sequence (SEQ ID NO. 172).

FIGS. 69A and 69B depict the nucleic acid sequence of the OspC-B31 (bp 55-633)/OspA-B31 (bp 394-651)/OspA-PKo (bp 652-820) chimer (SEQ ID NO. 173) and the encoded chimeric protein sequence (SEQ ID NO. 174).

FIGS. 70A and 70B depict the nucleic acid sequence of the OspC-B31 (bp 55-633)/OspA-K48 (bp 394-654)/OspA-Tro (bp 655-819) chimer (SEQ ID NO. 175) and the encoded chimeric protein sequence (SEQ ID NO. 176).

FIGS. 71A, 71B and 71C depict the nucleic acid sequence of the OspC-B31 (bp 55-633)/OspA-B31 (bp 88-492)/OspA-PKo (bp 493-537)/OspA-B31 (bp 541-651)/OspA-PKo (bp 652-822) chimer (SEQ ID NO. 177) and the encoded chimeric protein sequence (SEQ ID NO. 178); a variant of this sequence was also generated, where the N at position 190 of B31 OspA was deleted.

FIGS. 72A, 72B and 72C depict the nucleic acid sequence of the OspC-C2 (bp 55-612)/OspA-B31 (bp 88-492)/OspA-PKo (bp 493-537)/OspA-B31 (bp 541-651)/OspA-PKo (bp 652-822) chimer (SEQ ID NO. 179) and the encoded chimeric protein sequence (SEQ ID NO. 180); a variant of this sequence was also generated, where the N at position 190 of B31 OspA was deleted.

DETAILED DESCRIPTION OF THE INVENTION

The present invention pertains to chimeric proteins comprising various antigenic Borrelia polypeptides. In a preferred embodiment, the chimeric protein comprises Borrelia outer surface protein C (OspC) and outer surface protein A (OspA). These chimeric proteins have the general structure of OspC linked to OspA via a peptide bond. Each of the OspA and OspC portions of the chimeric OspC/OspA protein can be lipidated or unlipidated. In a preferred embodiment, the OspC/OspA chimer comprises OspC and OspA polypeptide fragments that do not possess their lipidation signals.

The chimeric forms of the OspA and OspC proteins described herein were bioengineered such that the resultant chimeric protein maintained at least some antigenicity of one or both of the parent molecules. As described herein, antigenic refers to the ability of a compound to bind products of an immune response, such as antibodies, T-cell receptors or both. Such responses can be measured using standard antibody detection assays, such as ELISA or standard T-cell activation assays. In a particular embodiment, the chimeric OspC/OspA proteins comprise OspA polypeptides which lack the putative autoreactive domain that has similarity to a region of human leukocyte function-associated antigen-1 (hLFA-1) (Gross, D. M. et al., Science, 281: 703-706 (1998)).

The current invention pertains to chimeric proteins comprising antigenic Borrelia polypeptides which do not occur in nature in the same Borrelia protein. The chimeric proteins are a combination of two or more antigenic polypeptides derived from Borrelia proteins. The antigenic polypeptides can be derived from different proteins from the same species of Borrelia, or different proteins from different Borrelia species, as well as from corresponding proteins from different species. As used herein, the term "chimeric protein" describes a protein comprising two or more polypeptides which are derived from corresponding and/or non-corresponding native Borrelia protein. A polypeptide "derived from" a native Borrelia protein is a polypeptide which has an amino acid sequence the same as an amino acid sequence present in a Borrelia protein, an amino acid sequence equivalent to the amino acid sequence of a naturally occurring Borrelia protein, or an amino acid sequence substantially similar to the amino acid sequence of a naturally occurring Borrelia protein (e.g., differing by a few amino acids), such as when a nucleic acid encoding a protein is subjected to site-directed mutagenesis. "Corresponding" proteins are equivalent proteins from different species or strains of Borrelia, such as outer surface protein A (OspA) from strain B31 and OspA from strain K48. The invention additionally pertains to nucleic acids encoding these chimeric proteins.

In one embodiment, the present invention is drawn to chimeric proteins comprising antigenic polypeptides from Lyme Disease-causing strains of Borrelia. In another embodiment, the chimeric proteins described herein comprise antigenic polypeptides from different pathogenic genospecies of Borrelia, such as Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii. In a preferred embodiment, the chimeric proteins comprise antigenic polypeptides from each of the pathogenic genospecies of Borrelia, including Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii.

The OspA portion of the chimeric molecules of the present invention can themselves be chimeric combinations of more than one OspA polypeptide. Similarly, the OspC portion of the chimeric molecules of the present invention can themselves be chimeric combinations of more than one OspC polypeptide. As described below, Applicants have identified two separate antigenic domains of OspA and OspB which flank the sole conserved tryptophan present in OspA and in OspB. These domains share cross-reactivity with different genospecies of Borrelia. The precise amino acids responsible for antigenic variability were determined through site-directed mutagenesis, so that proteins with specific amino acid substitutions are available for the development of chimeric versions of OspA which can be included in the OspC/OspA chimeric proteins of the present invention. Furthermore, Applicants have identified immunologically important hypervariable domains in OspA proteins, as described below in Example 2. The first hypervariable domain of interest for chimeric proteins, Domain A, includes amino acid residues 120-140 of OspA, the second hypervariable domain, Domain B, includes residues 150-180 and the third hypervariable domain, Domain C, includes residues 200-216 or 217 (depending on the position of the sole conserved tryptophan residue in the OspA of that particular species of Borrelia) (see FIG. 3). In addition, Applicants have sequenced the genes for several Borrelia proteins.

These discoveries have aided in the development of novel recombinant Borrelia proteins which include two or more amino acid regions or sequences which do not occur in the same Borrelia protein in nature. The recombinant proteins comprise polypeptides from a variety of Borrelia proteins, including, but not limited to, OspA, OspB, OspC, OspD, p12, p39, p41, p66, and p93. Preferred combinations include all or a portion of OspC linked to all or a portion of OspA. Antigenically relevant polypeptides from each of a number of proteins are combined into a single chimeric protein.

In one embodiment of the current invention, chimeras are now available which include antigenic OspA polypeptides flanking a tryptophan residue. OspB has a similar primary structure as OspA and is included in the following discussion. The antigenic polypeptides are derived from either the proximal portion from the tryptophan (the portion of the OspA protein present between the amino terminus and the conserved tryptophan of the protein), or the distal portion from the tryptophan (the portion of the OspA protein present between the conserved tryptophan of the protein and the carboxy terminus) in OspA. The resultant chimeras can be OspA-OspA chimeras (e.g., chimeras incorporating polypeptides derived from OspA from different strains of Borrelia), OspA-OspB chimeras, or OspB-OspB chimeras, and are constructed such that amino acid residues amino-proximal to an invariant tryptophan are from one protein and residues carboxy-proximal to the invariant tryptophan are from the other protein. For example, one available chimer consists of a polypeptide derived from the amino-proximal region of OspA from strain B31, followed by the tryptophan residue, followed by a polypeptide derived from the carboxy-proximal region of OspA from strain K48 (SEQ ID NO. 92). Another available chimer includes a polypeptide derived from the amino-proximal region of OspA from strain B31, and a polypeptide derived from the carboxy-proximal region of OspB from strain B31 (SEQ ID NO. 104). If the polypeptide proximal to the tryptophan of these chimeric proteins is derived from OspA, the proximal polypeptide can be further subdivided into the three hypervariable domains (Domains A, B, and C), each of which can be derived from OspA from a different strain of Borrelia. These chimeric proteins can further comprise antigenic polypeptides from another protein, e.g., OspC, in addition to the antigenic polypeptides flanking the tryptophan residue.

In one embodiment, the chimeric OspC/OspA proteins of the present invention comprise at least a first and a second polypeptide, wherein the first polypeptide comprises Borrelia burgdorferi OspC and wherein the second polypeptide comprises Borrelia burgdorferi OspA, such that OspC comprises the N-terminus of the protein.

In a particular embodiment, the first polypeptide comprises a Borrelia burgdorferi OspC polypeptide from about amino acid residue 19 to about amino acid residue 213, and the second polypeptide comprises a Borrelia burgdorferi OspA polypeptide. In another embodiment, the first polypeptide comprises a Borrelia burgdorferi OspC polypeptide from about amino acid residue 19 to about amino acid residue 211. In another embodiment, the first polypeptide comprises a Borrelia burgdorferi OspC polypeptide from about amino acid residue 19 to about amino acid residue 208. In another embodiment, the first polypeptide comprises a Borrelia burgdorferi OspC polypeptide from about amino acid residue 19 to about amino acid residue 204. The numbering of the OspC residues is according to the numbering of SEQ ID NO: 30 (FIGS. 12A and 12B). It is evident that the person of skill in the art recognizes that OspC genes from different strains and/or genospecies may differ in their primary sequence and that based on homology, similar regions of such OspC proteins could be identified and used in the present invention with no or only routine experimentation.

In one embodiment, the invention is drawn to chimeric OspC/OspA proteins wherein the first polypeptide comprises a Borrelia burgdorferi OspC polypeptide and the second polypeptide comprises a Borrelia burgdorferi OspA polypeptide from about amino acid residue 18 to about amino acid residue 273. In other embodiments, the chimeric OspC/OspA protein comprises a first polypeptide which is a Borrelia burgdorferi OspC polypeptide and a second polypeptide which is a Borrelia burgdorferi OspA polypeptide selected from the group consisting of an OspA polypeptide from about amino acid residue 132 to about amino acid residue 216, an OspA polypeptide from about amino acid residue 218 to about amino acid residue 273, an OspA polypeptide from about amino acid residue 18 to about amino acid residue 216 and an OspA polypeptide from about 132 to about amino acid residue 273. The numbering of the OspA residues is according to the numbering of SEQ ID NO: 7 (FIGS. 7A and 7B). It is evident that the person of skill in the art recognizes that OspA genes from different strains and/or genospecies may differ in their primary sequence and that based on homology, similar regions of such OspA proteins could be identified and used in the present invention with no or only routine experimentation.

The present invention is also drawn to OspC/OspA chimeric proteins wherein the first polypeptide comprises a Borrelia burgdorferi OspC polypeptide and the second polypeptide comprises a Borrelia burgdorferi OspA polypeptide, wherein the OspA polypeptide comprises two or more OspA polypeptide fragments as described above. In a preferred embodiment, the OspA polypeptide comprises portions of OspA from two or more strains of Borrelia. In another preferred embodiment, the OspA polypeptide comprises portions of OspA from two or more Lyme Disease-causing genospecies of Borrelia, e.g., wherein the genospecies are Borrelia burgdorferi sensu stricto, Borrelia afzelii and/or Borrelia garinii. In still another preferred embodiment, the OspC/OspA chimeric protein comprises one or more polypeptides from each of the pathogenic genospecies, Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii.

The chimeras described herein can be produced so that they are highly soluble, hyper-produced in E. coli, and non-lipidated. Lipidated chimeric proteins can also be produced. In addition, the chimeric proteins can be designed to end in an affinity tag (His-tag) to facilitate purification. The recombinant proteins described herein have been constructed to maintain antigenicity of at least one of the parent polypeptides. In addition, recombinant proteins specific for the various genospecies of Borrelia that cause Lyme disease are now available, because the genes from each of the major genospecies have been sequenced. These recombinant proteins with their novel biophysical and antigenic properties will be important diagnostic reagent and vaccine candidates.

The chimeric proteins of the current invention are advantageous in that they retain at least some specific reactivity to monoclonal or polyclonal antibodies that recognize wild-type Borrelia proteins. The proteins are immunogenic, and elicit antibodies that inhibit growth and/or induce lysis of Borrelia in vitro. Furthermore, in some embodiments, the proteins provide antigenic domains of two or more Borrelia strains and/or proteins within a single protein. Such proteins are particularly useful in immunodiagostic assays. For example, proteins of the present invention can be used as reagents in assays to detect the presence of antibodies to native Borrelia in potentially infected individuals. These proteins can also be used as immunodiagnostic reagents, such as in dot blots, Western blots, enzyme-linked immunosorbent assays (ELISA), or agglutination assays. The chimeric proteins of the present invention can be produced by known techniques, such as by recombinant methodology, polymerase chain reaction, or mutagenesis.

Furthermore, the proteins of the current invention are useful as vaccine immunogens against Borrelia infection. Because Borrelia has been shown to be clonal, a protein comprising antigenic polypeptides from a variety of Borrelia proteins and/or species, will provide immunoprotection for a considerable time when used in a vaccine. The lack of significant intragenic recombination, a process which might rapidly generate novel epitopes with changed antigenic properties, ensures that Borrelia can only change antigenic type by accumulating mutational change, which is slow when compared with recombination in generating different antigenic types. The chimeric protein can be combined with a physiologically-acceptable carrier and administered to a vertebrate animal through standard methods (e.g., intravenously or intramuscularly, for example).

In addition to the chimeric proteins described herein, the present invention is also drawn to nucleic acids which encode the Borrelia chimeric protein described herein. In one embodiment of the present invention, the composition comprises a nucleic acid that encodes a chimeric protein of at least two polypeptides, wherein the first polypeptide comprises Borrelia burgdorferi OspC, and the second polypeptide comprises Borrelia burgdorferi OspA, such that OspC is upstream of OspA. The OspC and OspA nucleic acid fragments that make up the chimeric protein can be from the same strain or genospecies of Borrelia, from different strains or genospecies of Borrelia, or from combinations of nucleic acids that are from the same and/or different strains or genospecies of Borrelia.

It is understood that the nucleic acids that encode the polypeptides that comprise the chimeric protein can include extra nucleotides or fewer nucleotides in order to simplify the construction of the gene encoding the chimeric polypeptide, e.g., to allow for the use of convenient restriction endonuclease sites or to allow the ligation of the gene fragments such that a contiguous coding region is created. Based on the guidance provided herein, one of ordinary skill in the art would readily be able to add or remove nucleotides from the termini of the gene fragments encoding the polypeptides of the chimeric OspC/OspA protein in order to generate the chimeric proteins of the present invention with no or only routine experimentation. Furthermore, there can be an extra about 1 to about 10 amino acids on the N- and/or C-terminus of the polypeptides and chimeric proteins of the present invention and still retain the properties of the present invention. It is also understood that those of skill in the art, using art-known methods and/or the methods described herein, could generate additional OspC-OspA chimeric proteins, and that these chimeric proteins are encompassed by the invention.

The present invention is also drawn to expression vectors which comprise an isolated DNA encoding the Borrelia chimeric protein described herein. In one embodiment, the composition includes an expression vector comprising an isolated DNA which encodes an OspC/OspA chimeric protein, wherein the OspC portion of the protein is upstream of the OspA portion. The present invention also encompasses host cells which comprise a recombinant nucleic acid that encodes an OspC/OspA chimeric protein, as described herein.

The present invention is also drawn to methods of making the Borrelia chimeric polypeptides described herein. In one embodiment, the method of making a chimeric Borrelia protein comprises selecting a polynucleotide sequence encoding OspC, or an antigenic portion thereof, selecting a polynucleotide sequence encoding OspA, or an antigenic portion thereof, and ligating these polynucleotide sequences together, such that OspC comprises the N-terminus of the protein. The polypeptides of the present invention can also be recombinantly expressed in suitable microbial hosts, wherein said hosts include, but are not limited to, bacterial hosts, such as E. coli, fungal hosts, such as S. cerevisiae or cell culture hosts, such as those of mammalian cell culture or insect cell culture.

The present invention is also drawn to methods of delivering the Borrelia chimeric polypeptides described herein. In one embodiment, the method comprises administering the chimeric protein in a physiologically-acceptable carrier to an individual. The individual develops at least some immune response to the chimeric protein. As an example, the individual could generate a humoral immune response, wherein antibodies that recognize at least a portion of said chimeric polypeptide are produced by the individual. The antibodies that recognize the chimeric polypeptide can be of any class of immunoglobulin, such as IgM, IgD, IgA and IgG or combinations thereof.

The present invention is also drawn to methods of delivering a nucleic acid which encodes a chimeric polypeptide described herein. In one embodiment, the method comprises administering the nucleic acid in a physiologically-acceptable carrier to an individual using art-accepted methods of DNA delivery, including but not limited to, biolistic delivery and lipid encapsulation. The chimeric polypeptide is at least transiently expressed and the individual develops at least some immune response to the chimeric protein encoded by the nucleic acid.

The invention also encompasses methods of using the chimeric proteins described herein in diagnostic assays. In one embodiment, the method can be used to detect the presence of OspA- and/or OspC-specific antibodies in a sample, e.g., a host sample of interest. In one embodiment, the method comprises contacting a host sample of interest with the chimeric OspC/OspA protein, under conditions, wherein antibodies, if present in the host sample, bind to the chimeric protein thereby forming antigen-antibody complexes. The antigen-antibody complexes are then detected. In this manner, an immune response to Lyme-Disease causing Borrelia can be detected.

As described herein, the chimeric proteins of the present invention incorporate antigenic domains from different Borrelia proteins, as well as from different Borrelia strains and/or genospecies. As such, they are useful in the detection or diagnosis of the presence of Lyme disease-causing Borrelia, especially Borrelia from groups capable of causing disseminated symptoms of Lyme disease. Disseminated symptoms refer to infection outside of the erythema migrans skin lesion, e.g., infection in blood, CNS or synovia.

The chimeric polypeptides of the present invention elicit specific immune responses to OspC and OspA. In one embodiment, the chimeric polypeptides elicit immune responses to strains of Lyme disease-causing Borrelia of the same genospecies as that represented by the OspC/OspA chimeric protein. In another embodiment, the chimeric polypeptides elicit immune responses to strains of Lyme disease-causing Borrelia of different genospecies than that represented by the OspC/OspA chimeric protein, as well as to Lyme disease-causing Borrelia of the same genospecies as that represented by the OspC/OspA chimeric protein. The immune response includes, but is not limited to, a humoral response, a secretory response, a cell-mediated response, or any combination thereof.

The immunogenic compositions of the present invention can also be used to immunize animals, e.g., mammals, including humans. Immunization is understood to elicit specific immunogenic responses as described herein. In one embodiment, administration of an immunogenic composition, e.g., an OspC/OspA chimeric protein, an OspC/OspA chimeric nucleic acid, to an animal results in the animal developing immunity to infection by Lyme disease-causing Borrelia, e.g., Borrelia burgdorferi, Borrelia afzelii and/or Borrelia garinii.

Immunity, as described herein, is understood to mean the ability of the treated animal to resist infection (e.g., systemic infection), to overcome infection (e.g., systemic infection) or to overcome infection (e.g., systemic infection) more easily and/or more quickly when compared to non-immunized and/or non-treated individuals. Immunity can also include an improved ability of the treated individual to sustain an infection with reduced or no clinical symptoms of systemic infection. The individual may be treated with the chimeric proteins of the present invention either proactively, e.g., once a year or alternatively after sustaining a tick bite.

In one embodiment, the OspC/OspA chimeric protein of the present invention, together with suitable excipients and/or adjuvants, is administered to an animal such that the animal develops an immune response to at least one of the OspC and/or OspA polypeptides of the composition. The pharmaceutical composition can also be administered with other components suitable for in vitro and/or in vivo use. These additional components include buffers, carrier proteins, adjuvants, excipients, preservatives and combinations thereof.

The present invention is also drawn to pharmaceutical compositions which can be used to vaccinate and/or treat Borrelia infection in an animal or human. In a particular embodiment, the pharmaceutical composition comprises a Borrelia burgdorferi OspC/OspA chimeric protein. The pharmaceutical composition can also be administered together with a physiologically-acceptable carrier, an excipient and/or an adjuvant. Suitable adjuvants are well known in the art (see for example PCT Publication WO 96/40290, the entire teachings of which are incorporated herein by reference), and can be used, for example, to enhance immunogenicity, potency or half-life of the chimeric proteins in the treated animal.

The pharmaceutical compositions used to vaccinate and/or treat Borrelia infection can be prepared using methods for preparing vaccines which are well known in the art. For example, the OspC/OspA chimeric proteins described herein can be isolated and/or purified by known techniques, such as by size exclusion chromatography, affinity chromatography, ion exchange chromatography, preparative electrophoresis, selective precipitation or combinations thereof. The prepared chimeric proteins can be mixed with suitable other reagents as described herein, such that the chimeric protein is at a suitable concentration. The dosage of the chimeric protein will vary and depends upon the age, weight and/or physical condition of the animal, e.g., mammal, human, to be treated. The optimal dosage can be determined by routine optimization techniques, using suitable animal models.

Administration of the pharmaceutical composition to be used as a vaccine can be by any suitable technique. Suitable techniques for administration of the pharmaceutical composition include, but are not limited to, injection, e.g., subcutaneous injection, intramuscular injection, intravenous injection, intra peritoneal injection; mucosal administration, e.g., exposing nasal mucosa to nose drops containing the proteins or chimeric proteins of the present invention; oral administration; and DNA immunization.

The incorporation of polypeptide fragments from different strains and/or genospecies of Borrelia burgdorferi allows for a greater detection range and a more effective vaccination tool. The present invention provides a chimeric combination of proteins that, when used as a vaccine, can prevent Lyme disease from becoming systemic. The chimeric proteins of the present invention can be effective in preventing Lyme disease, as well as having a therapeutic effect on established infection, for example, after the tick bite is noticed by the patient. Since the chimeric proteins of the present invention comprise both OspC and OspA polypeptides, they are expected to act at the level of the tick as well as the level of the host in preventing both infection and disease due to Borrelia burgdorferi, Borrelia afzelii and/or Borrelia garinii.

The present invention is also drawn to diagnostic kits which comprise the chimeric polypeptides described herein. In one embodiment, the kit comprises a chimeric protein comprising at least a first and a second polypeptide, wherein the first polypeptide comprises Borrelia burgdorferi OspC and wherein the second polypeptide comprises Borrelia burgdorferi OspA, such that OspC comprises the N-terminus of the protein. The kit also includes reagents for detecting antibody-antigen complexes that are formed between the OspC/OspA chimeric protein and antibodies that are present in a sample, e.g., a user-supplied host sample.

As a result of the present invention, it is now possible to prepare improved diagnostic tools comprising both OspA and OspC antigens from various Borrelia burgdorferi strains and/or genospecies. Since OspA is primarily expressed in the tick vector, and OspC is upregulated in response to the feeding of an infected tick on a mammal, the diagnostic compositions of the invention can recognize antigens that are expressed at different stages of the life cycle of Borrelia burgdorferi. Moreover, by incorporating unique polypeptide fragments from pathogenic families of Borrelia, the present invention allows for improved diagnostic compositions which can detect clinically important exposure to pathogenic Borrelia while overlooking the remainder of non-pathogenic families of Borrelia.

As described herein, the OspC/OspA chimeric proteins were bioengineered such that the protective domains of each protein were maintained. In experiments described herein, mice were either immunized with OspA, OspC or OspC/OspA chimeric proteins in aluminum hydroxide. Mice were then bled and tested for antibody responses against OspC and OspA derived from various strains of Borrelia. In additional experiments, these immunized mice were challenged with ticks infected with Borrelia burgdorferi and transmission of infection was assessed.

Mice immunized with the OspC/OspA chimeric protein gave a remarkable and equivalent antibody response to both OspA and OspC, as compared to mice immunized with OspA and OspC control proteins (FIGS. 47 and 48). In addition, antibodies in the sera of mice immunized with the OspC/OspA chimeric protein were also reactive against antigens derived from different strains of Borrelia burgdorferi (FIGS. 49-50 and 52-54). Chimer-immunized mice were fully protected against challenge with ticks infected with Borrelia burgdorferi, as compared to sham-vaccinated controls (infection rates of 100%) (Table VI).

In other experiments described herein, mice were either immunized with a lipidated OspA chimeric protein, a lipidated OspC chimeric protein, or a non-lipidated OspC/OspA chimeric protein, once again in the presence of aluminum hydroxide. Mice were then bled and tested for antibody responses against OspA and OspC derived from various strains of Borrelia. Surprisingly, the results of these studies indicate that mice immunized with the non-lipidated OspC/OspA chimeric protein have antibody responses to OspA and OspC that are equivalent or greater than those generated by mice immunized with the corresponding lipidated OspA or lipidated OspC chimeric proteins (FIGS. 49-51).

The results of the studies presented herein indicate that mice immunized with OspC-OspA chimeric proteins generate a potent antibody response against two immunoprotective targets that are expressed at different stages of the life cycle of Borrelia burgdorferi.

The current invention is illustrated by the following Examples, which are not to be construed to be limiting in any way.

EXEMPLIFICATION

Example 1

Purification of Borrelia burgdorferi Outer Surface Protein A and Analysis of Antibody Binding Domains

This example details a method for the purification of large amounts of native outer surface protein A (OspA) to homogeneity, and describes mapping of the antigenic specificities of several anti-OspA MAbs. OspA was purified to homogeneity by exploiting its resistance to trypsin digestion. Intrinsic labeling with .sup.14C-palmitic acid confirmed that OspA was lipidated, and partial digestion established lipidation at the amino-terminal cysteine of the molecule.

The reactivity of seven anti-OspA murine monoclonal antibodies to nine different Borrelia isolates was ascertained by Western blot analysis. Purified OspA was fragmented by enzymatic or chemical cleavage, and the monoclonal antibodies were able to define four distinct immunogenic domains (see FIG. 1). Domain 3, which included residues 190-220 of OspA, was reactive with protective antibodies known to agglutinate the organism in vitro, and included distinct specificities, some of which were not restricted to a genotype of B. burgdorferi.

A. Purification of Native OspA

Detergent solubilization of B. burgdorferi strips the outer surface proteins and yields partially-purified preparations containing both OspA and outer surface protein B (OspB) (Barbour, A. G. et al., Infect. Immun. 52 (5): 549-554 (1986); Coleman, J. L. and J. L. Benach, J Infect. Dis. 155 (4): 756-765 (1987); Cunningham, T. M. et al., Ann. NY Acad. Sci. 539: 376-378 (1988); Brandt, M. E. et al., Infect. Immun. 58: 983-991 (1990); Sambri, V. and R. Cevenini, Microbiol. 14: 307-314 (1991)). Although both OspA and OspB are sensitive to proteinase K digestion, in contrast to OspB, OspA is resistant to cleavage by trypsin (Dunn, J. et al., Prot. Exp. Purif. 1: 159-168 (1990); Barbour, A. G. et al., Infect. Immun. 45: 94-100 (1984)). The relative insensitivity to trypsin is surprising in view of the fact that OspB A has a high (16% for B31) lysine content, and may relate to the relative configuration of OspB A and B in the outer membrane.

Intrinsic Radiolabeling of Borrelia

Labeling for lipoproteins was performed as described by Brandt et al. (Brandt et al., Infect. Immun. 58: 983-991 (1990)). .sup.14C-palmitic acid (ICN, Irvine, Calif.) was added to the BSK II media to a final concentration of 0.5 .mu.Ci per milliliter (ml). Organisms were cultured at 34.degree. C. in this medium until a density of 10.sup.8 cells per ml was achieved.

Purification of OspA Protein from Borrelia Strain B31

Borrelia burgdorferi, either .sup.14C-palmitic acid-labeled or unlabeled, were harvested and washed as described (Brandt, M. E. et al., Infect. Immun. 58: 983-991 (1990)). Whole organisms were trypsinized according to the protocol of Barbour et al. (Infect. Immun. 45: 94-100 (1984)) with some modifications. The pellet was suspended in phosphate buffered saline (PBS, 10 mM, pH 7.2), containing 0.8% tosyl-L-phenylalanine chloromethyl ketone (TPCK)-treated trypsin (Sigma, St. Louis, Mo.), the latter at a ratio of 1 .mu.g per 10.sup.8 cells. Reaction was carried out at 25.degree. C. for 1 hour, following which the cells were centrifuged. The pellet was washed in PBS with 100 .mu.g/ml phenylmethylsulfonyl fluoride (PMSF). Triton X-114 partitioning of the pellet was carried out as described by Brandt et al. (Brandt et al., Infect. Immun. 58: 983-991 (1990)). Following trypsin treatment, cells were resuspended in ice-cold 2% (v/v) Triton X-114 in PBS at 10.sup.9 cells per ml. The suspension was rotated overnight at 4.degree. C., and the insoluble fraction removed as a pellet after centrifugation at 10,000.times.g for 15 minutes at 4.degree. C. The supernatant (soluble fraction) was incubated at 37.degree. C. for 15 minutes and centrifuged at room temperature at 1000.times.g for 15 minutes to separate the aqueous and detergent phases. The aqueous phase was decanted, and ice cold PBS added to the lower Triton phase, mixed, warmed to 37.degree. C., and again centrifuged at 1000.times.g for 15 minutes. Washing was repeated twice more. Finally, detergent was removed from the preparation using a spin column of Bio-beads SM2 (BioRad, Melville, N.Y.) as described (Holloway, P. W., Anal. Biochem. 53: 304-308 (1973)).

Ion exchange chromatography was carried out as described by Dunn et al. (Dunn et al., Prot. Exp. Purif. 1: 159-168 (1990)) with minor modifications. Crude OspA was dissolved in buffer A (1% Triton X-100, 10 mM phosphate buffer (pH 5.0)) and loaded onto a SP Sepharose resin (Pharmacia, Piscataway, N.J.), pre-equilibrated with buffer A at 25.degree. C. After washing the column with 10 bed-volumes of buffer A, the bound OspA was eluted with buffer B (1% Triton X-100, 10 mM phosphate buffer (pH 8.0)). OspA fractions were detected by protein assay using the BCA method (Pierce, Rockford, Ill.), or as radioactivity when intrinsically labeled material was fractionated. Triton X-100 was removed using a spin column of Bio-beads SM2.

This method purifies OspA from an outer surface membrane preparation. In the absence of trypsin-treatment, OspA and B were the major components of the soluble fraction obtained after Triton partitioning of strain B31. In contrast, when Triton extraction was carried out after trypsin-treatment, the OspB band is not seen. Further purification of OspA-B31 on a SP Sepharose column resulted in a single band by SDS-PAGE. The yield following removal of detergent was approximately 2 mg per liter of culture. This method of purification of OspA, as described herein for strain B31, can be used for other isolates of Borrelia as well. For strains such as strain K48, which lack OspB, trypsin treatment can be omitted.

Lipidation Site of OspA-B31

.sup.14C-palmitic acid labeled OspA from strain B31 was purified as described above and partially digested with endoproteinase Asp-N (data not shown). Following digestion, a new band of lower molecular weight was apparent by SDS-PAGE, found by direct amino-terminal sequencing to begin at Asp.sub.25. This band had no trace of radioactivity by autoradiography (data not shown). OspA and B contain a signal sequence (L-X--Y--C) similar to the consensus described for lipoproteins of E. coli, and it has been predicted that the lipidation site of OspA and B should be the amino-terminal cysteine (Brandt, M. E. et al., Infect. Immun 58: 983-991 (1990)). The results presented herein support this prediction.

B. Comparison of OspA Antibody Binding Regions in Nine Strains of Borrelia burgdorferi

The availability of the amino acid sequenced for OspA from a number of different isolates, combined with peptide mapping and Western blot analysis, permitted the identification of the antigenic domains recognized by monoclonal antibodies (MAbs) and allowed inference of the key amino acid residues responsible for specific antibody reactivity.

Strains of Borrelia burgdorferi

Nine strains of Borrelia, including seven European strains and two North American strains, were used in this study of antibody binding domains of several proteins. Information concerning the strains is summarized in Table I, below.

TABLE-US-00001 TABLE I Representative Borrelia Strains Strain Location and Source Reference for Strain K48 Czechoslovakia, none Ixodes ricinus PGAU Germany, human ACA Wilske, B. et al., J. Clin. Microbiol 32: 340-350 (1993) DK29 Denmark, human EM Wilske, B. et al. PKo Germany, human EM Wilske, B. et al. PTrob Germany, human skin Wilske, B. et al. Ip3 Khabarovsk, Russia, Asbrink, E. et al., Acta I. persulcatus Derm. Venereol. 64: 506-512 (1984) Ip90 Khabarovsk, Russia, Asbrink, E. et al. I. persulcatus 25015 Millbrook, NY, Barbour, A. G. et al., Curr. I. persulcatus Microbiol. 8: 123-126 (1983) B31 Shelter Island, NY, Luft, B. J. et al., Infect. I. scapularis Immun. 60: 4309-4321 (1992); ATCC 35210 PKa1 Germany, human CSF Wilske, B. et al. ZS7 Freiburg, Germany, Wallich, R. et al., Nucl. I. ricinus Acids Res. 17: 8864 (1989) N40 Westchester Co., NY Fikrig, E. et al., Science 250: 553-556(1990) PHei Germany, human CSF Wilske, B. et al. ACAI Sweden, human ACA Luft, B. J. et al., FEMS Microbiol. Lett. 93: 73-68 (1992) PBo Germany, human CSF Wilske, B. et al. ACA = patient with acrodermatitis chronica atrophicans; EM = patient with erythema migrans; CSF = cerebrospinal fluid of patient with Lyme disease

Strains K48, PGAU and DK29 were supplied by R. Johnson, University of Minnesota; PKo and PTrob were provided by B. Wilske and V. Preac-Mursic of the Pettenkhofer Institute, Munich, Germany; and Ip3 and Ip90 were supplied by L. Mayer of the Center for Disease Control, Atlanta, Ga. The North American strains included strain 25015, provided by J. Anderson of the Connecticut Department of Agriculture; and strain B31 (ATCC 35210).

Monoclonal Antibodies

Seven monoclonal antibodies (MAbs) were utilized in this study. Five of the MAbs (12, 13, 15, 83 and 336) were produced from hybridomas cloned and subcloned as previously described (Schubach, W. H., et al., Infect. Immun. 59(6): 1911-1915 (1991)). MAb H5332 (Barbour, A. G. et al., Infect. Immun. 41: 795-804 (1983)) was a gift from Drs. Alan Barbour, University of Texas, and MAb CIII.78 (Sears, J. E. et al., J. Immunol. 147(6):1995-2000 (1991)) was a gift from Richard A. Flavell, Yale University. MAbs 12 and 15 were raised against whole sonicated B3; MAb 336 was produced against whole PGAU; and MAbs 13 and 83 were raised to a truncated form of OspA cloned from the K48 strain and expressed in E. coli using the T7 RNA polymerase system (McGrath, B. C. et al., Vaccines, Cold Spring Harbor Laboratory Press, Plainview, N.Y., pp. 365-370 (1993)). All MAbs were typed as being Immunoglobulin G (IgG).

Methods of Protein Cleavage, Western Blotting and Amino-Terminal Sequencing

Prediction of the various cleavage sites was achieved by knowledge of the primary amino acid sequence derived from the full nucleotide sequences of OspA, many of which are currently available (see Table II, below). Cleavage sites can also be predicted based on the peptide sequence of OspA, which can be determined by standard techniques after isolation and purification of OspA by the method described above. Cleavage of several OspA isolates was conducted to determine the localization of monoclonal antibody binding of the proteins.

Hydroxylamine-HCl (HA), N-chlorosuccinimide (NCS), and cyanogen bromide cleavage of OspA followed the methods described by Bornstein (Biochem. 9 (12):2408-2421 (1970)), Shechter et al., (Biochem. 15 (23):5071-5075 (1976)), and Gross (in Hirs, C. H. W. (ed): Methods in Enzymology, (N.Y. Acad. Press), 11:238-255 (1967)) respectively. Protease cleavage by endoproteinase, Asp-N (Boehringer Mannheim, Indianapolis, Ind.), was performed as described by Cleveland D. W. et al., (J. Biol. Chem. 252: 1102-1106 (1977)). Ten micrograms of OspA were used for each reaction. The ratio of enzyme to OspA was approximately 1 to 10 (w/w).

Proteins and peptides generated by cleavage were separated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) (Laemmli, U.K., Nature (London) 227:680-685 (1970)), and electroblotted onto immobilon Polyvinylidine Difluoride (PVDF) membranes (Ploskal, M. G. et al., Biotechniques 4: 272-283 (1986)). They were detected by amido black staining or by immunostaining with murine MAbs, followed by alkaline phosphatase-conjugated goat antimouse IgG. Specific binding was detected using a 5-bromo-4-chloro-3-indolylphosphate (BCIP)/nitroblue tetrazolium (NBT) developer system (KPL Inc., Gathersburg, Md.).

In addition, amino-terminal amino acid sequence analysis was carried out on several cleavage products, as described by Luft et al. (Infect. Immun. 57: 3637-3645 (1989)). Amido black stained bands were excised from PVDF blots and sequenced by Edman degradation using a Biosystems model 475A sequenator with model 120A PTH analyzer and model 900A control/data analyzer.

Cleavage Products of Outer Surface Protein A Isolates

Purified OspA-B31, labeled with .sup.14C-palmitic acid, was fragmented with hydroxylamine-HCl (HA) into two peptides, designated HA1 and HA2 (data not shown). The HA1 band migrated at 27 KD and retained its radioactivity, indicating that the peptide included the lipidation site at the N-terminus of the molecule (data not shown). From the predicted cleavage point, HA1 should correspond to residues 1 to 251 of OspA-B31. HA2 had a MW of 21.6 KD by SDS-PAGE, with amino-terminal sequence analysis showing it to begin at Gly72, i.e. residues 72 to 273 of OspA-B31. By contrast, HA cleaved OspA-K48 into three peptides, designated HA1, HA2, and HA3 with apparent MWs of 22 KD, 16 KD and 12 KD, respectively. Amino-terminal sequencing showed HA1 to start at Gly72, and HA3 at Gly142. HA2 was found to have a blocked amino-terminus, as was observed for the full-length OspA protein. HA1, 2 and 3 of OspA-K48 were predicted to be residues 72-274, 1 to 141 and 142 to 274, respectively.

N-Chlorosuccinimide (NCS) cleaves tryptophan (W), which is at residue 216 of OspA-B31 or residue 217 of OspA-K48 (data not shown). NCS cleaved OspA-B31 into 2 fragments, NCS1, with MW of 23 KD, residues 1-216 of the protein, and NCS2 with a MW of 6.2 KD, residues 217 to 273 (data not shown). Similarly, K48 OspA was divided into 2 pieces, NCS1 residues 1-217, and NCS2 residues 218 to 274 (data not shown).

Cleavage of OspA by cyanogen bromide (CNBr) occurs at the carboxy side of methionine, residue 39. The major fragment, CNBr1, has a MW of 25.7 KD, residues 39-274 by amino-terminal amino acid sequence analysis (data not shown). CNBr2 (about 4 KD) could not be visualized by amido black staining; instead, lightly stained bands of about 20 KD MW were seen. These bands reacted with anti-OspA MAbs, and most likely were degradation products due to cleavage by formic acid.

Determination of Antibody Binding Domains for Anti-OspA Monoclonal Antibodies

The cleavage products of OspA-B31 and OspA-K48 were analyzed by Western blot to assess their ability to bind to the six different MAbs. Preliminary Western blot analysis of the cleavage products demonstrated that strains K48 and DK29 have similar patterns of reactivity, as do Ip3, PGAU and PKo. The OspA of strain PTrob was immunologically distinct from the others, being recognized only by MAb 336. MAb 12 recognized only the two North American strains, B31 and 25015. When the isolates were separated into genogroups, it was remarkable that all the MAbs, except MAb 12, crossed over to react with multiple genogroups.

MAb12, specific for OspA-B31, bound to both HA1 and HA2 of OspA-B31. However, cleavage of OspA-B31 by NCS at residue Trp216 created fragments which did not react with MAb12, suggesting that the relevant domain is near or is structurally dependent upon the integrity of this residue (data not shown). MAb 13 bound only to OspA-K48, and to peptides containing the amino-terminus of that molecule (e.g. HA2; NCS1). It did not bind to CNBr1 residues 39 to 274. Thus the domain recognized by MAb13 is in the amino-terminal end of OspA-K48, near Met38.

MAb 15 reacts with the OspA of both the B31 and K48 strains, and to peptides containing the N-terminus of OspA, such as HA1 of OspA-B31 and NCS1, but not to peptides HA2 of OspA-B31 and HA1 of OspA-K48 (data not shown). Both peptides include residue 72 to the C-terminus of the molecules. MAb15 bound to CNBr1 of OspA-K48, indicating the domain for this antibody to be residues 39 to 72, specifically near Gly72 (data not shown).

MAb83 binds to OspA-K48, and to peptides containing the C-terminal portion of the molecule, such as HA1. They do not bind to HA2 of OspA-K48, most likely because the C-terminus of HA2 of OspA-K48 ends at 141. Similar to MAb12 and OspA-B31, binding of MAbs 83 and CIII.78 is eliminated by cleavage of OspA at the tryptophan residue. Thus binding of MAbs 12, 83 and CIII.78 to OspA depends on the structural integrity of the Trp.sub.216 residue, which appears to be critical for antigenicity. Also apparent is that, although these MAbs bind to a common antigenic domain, the precise epitopes which they recognize are distinct from one another given the varying degrees of cross-reactivity to these MAbs among strains.

Although there is similar loss of binding activity of MAb336 with cleavage at Trp.sub.216, this MAb does not bind to HA1 of OspA-B31, suggesting the domain for this antibody includes the carboxy-terminal end of the molecule, inclusive of residues 251 to 273. Low MW peptides, such as HA3 (10 KD) and NCS2 (6 KD), of OspA-K48 do not bind this MAb on Western blots. In order to confirm this observation, we tested binding of the 6 MAbs with a recombinant fusion construct p3A/EC that contains a trpE leader protein fused with residues 217 to 273 of OspA-B31 (Schubach, W. H. et al., Infect. Immun. 59(6): 1911-1915 (1991)). Only MAb336 reacted with this construct (data not shown). Peptides and antigenic domains localized by fragmentation of OspA are summarized in FIG. 1.

Mapping of Domains to Define the Molecular Basis for the Serotype Analysis

To define the molecular basis for the serotype analysis of OspA, we compared the derived amino acid sequences of OspA for the nine isolates (FIG. 2). At the amino terminus of the protein, these predictions can be more precise given the relatively small number of amino acid substitutions in this region compared to the carboxy terminus. Domain 1, which is recognized by MAb13, includes residues Leu34 to Leu41. MAb13 only binds to the OspA of species K48, DK29 and IP90. Within this region, residue 37 is variable, however Gly37 is conserved amongst the three reactive strains. When Gly37 is changed to Glu37, as it is in OspA of strains B31, PTrob, PGAU, and PKo, MAb13 does not recognize the protein (data not shown). By similar analysis, it can be seen that Asp70 is a crucial residue for Domain 2, which includes residues 65 to 75 and is recognized by MAb15. Domain 3 is reactive with MAbs H5332, 12 and 83, and includes residues 190-220. It is clear that significant heterogeneity exists between MAbs reactive with this domain, and that more than one conformational epitope must be contained within the sequence. Domain 4 binds MAb336, and includes residues 250 to 270. In this region, residue 266 is variable and therefore may be an important determinant. It is apparent, however, that other determinants of the reactivity of this monoclonal antibody reside in the region comprising amino acids 217-250. Furthermore, the structural integrity of Trp216 is essential for antibody reactivity in the intact protein. Finally, it is important to stress that FIG. 2 indicates only the locations of the domains, and does not necessarily encompass the entire domain. Exact epitopes are being analyzed by site-directed mutagenesis of specific residues.

Overall, evidence suggests that the N-terminal portion is not the immunodominant domain of OspA, possibly by virtue of its lipidation, and the putative function of the lipid moiety in anchoring the protein to the outer envelope. The C-terminal end is immunodominant and includes domains that account in part for structural heterogeneity (Wilske, B. et al., Med. Microbiol. Immunol. 181: 191-207 (1992)), and may provide epitopes for antibody neutralization (Sears, J. E. et al., J. Immunol. 147(6): 1995-2000 (1991)), and relate to other activities, such as the induction of T-cell proliferation (Shanafel, M. M., et al., J. Immunol. 148: 218-224 (1992)). There are common epitopes in the carboxy-end of the protein that are shared among genospecies which may have immunoprotective potential (Wilske, B., et al., Med. Microbiol. Immunol. 181: 191-207 (1992)).

Prediction of secondary structure on the basis of hydropathy analysis and circular dichroism and fluorescence spectroscopy measurements (McGrath, B. C., et al., Vaccines, Cold Spring Harbor Laboratory Press, Plainview, N.Y.; pp. 365-370 (1993)) suggest domains 3 and 4 to be in a region of the molecule with a propensity to form alpha-helix, whereas domains 1 and 2 occur in regions predicted to be beta-sheets (see FIG. 1). These differences may distinguish domains in accessibility to antibody or to reactive T-cells (Shanafel, M. M. et al., J. Immunol. 148: 218-224 (1992)). Site-directed mutagenesis of specific epitopes, as described below in Example 2, aids in identifying exact epitopes.

Example 2

Identification of an Immunologically Important Hypervariable Domain of the Major Outer Surface Protein A of Borrelia

This Example describes epitope mapping studies using chemically cleaved OspA and TrpE-OspA fusion proteins. The studies indicate a hypervariable region surrounding the single conserved tryptophan residue of OspA (at residue 216, or in some cases 217), as determined by a moving window population analysis of OspA from fifteen European and North American isolates of Borrelia. The hypervariable region is important for immune recognition.

Site-directed mutagenesis was also conducted to examine the hypervariable regions more closely. Fluorescence and circular dichroism spectroscopy have indicated that the conserved tryptophan is part of an alpha-helical region in which the tryptophan is buried in a hydrophobic environment (McGrath, B. C., et al., Vaccines, Cold Spring Harbor Laboratory Press, Plainview, N.Y.; pp. 365-370 (1993)). More polar amino acid side-chains flanking the tryptophan are likely to be exposed to the hydrophilic solvent. The hypervariability of these solvent-exposed residues among the various strains of Borrelia suggested that these amino acid residues may contribute to the antigenic variation in OspA. Therefore, site-directed mutagenesis was performed to replace some of the potentially exposed amino acid side chains in the protein from one strain with the analogous residues of a second strain. The altered proteins were then analyzed by Western Blot using monoclonal antibodies which bind OspA on the surface of the intact, non-mutated spirochete. The results indicated that certain specific amino acid changes near the tryptophan can abolish reactivity of OspA to these monoclonal antibodies.

A. Verification of Clustered Polymorphisms in Outer Surface Protein A Sequences

Cloning and sequencing of the OspA protein from fifteen European and North American isolates (described above in Table I) demonstrated that amino acid polymorphism is not randomly distributed throughout the protein; rather, polymorphism tended to be clustered in three regions of OspA. The analysis was carried out by plotting the moving, weighted average polymorphism of a window (a fixed length subsection of the total sequence) as it is slid along the sequence. The window size in this analysis was thirteen amino acids, based upon the determination of the largest number of significantly deviating points as established by the method of Tajima (J. Mol. Evol. 33: 470-473 (1991)). The average weighted polymorphism was calculated by summing the number of variant alleles for each site. Polymorphism calculations were weighted by the severity of amino acid replacement (Dayhoff, M. O. et al., in: Dayhoff, M. O. (ed.) Atlas of protein Sequence and Structure NBRF, Washington, Vol. 5, Suppl. 3: 345 (1978)). The sum was normalized by the window size and plotted. The amino acid sequence position corresponds to a window that encompasses amino acids 1 through 13. Bootstrap resampling was used to generate 95% confidence intervals on the sliding window analysis. Since Borrelia has been shown to be clonal, the bootstrap analysis should give a reliable estimate of the expected variance from polymorphism calculations. The bootstrap was iterated five hundred times at each position, and the mean was calculated from the sum of all positions. The clonal nature of Borrelia ensures that the stochastic variance that results from differing genealogical histories of the sequence positions (as would be expected if recombination were prevalent) will be minimized.

This test verified that the three regions around the observed peaks all have significant excesses of polymorphism. Excesses of polymorphism were observed in the regions including amino acid residues 132-145, residues 163-177, and residues 208-221 (FIG. 3). An amino acid alignment between residues 200 and 220 for B31, K48 and the four site-directed mutants is shown in FIG. 4. The amino acid 208-221 region includes the region of OspA which has been modeled as an oriented alpha-helix in which the single tryptophan residue at amino acid 216 is buried in a hydrophobic pocket, thereby exposing more polar amino acids to the solvent (FIG. 5) (France, L. L., et al., Biochem. Biophys. Acta 1120: 59 (1992)). These potentially solvent-exposed residues showed considerable variability among the OspAs from various strains and may be an important component of OspA antigenic variation. For the purposes of generating chimeric proteins, the hypervariable domains of interest are Domain A, which includes amino acid residues 120-140 of OspA; Domain B, which includes residues 150-180; and Domain C, which includes residues 200-216 or 217.

B. Site-Directed Mutagenesis of the Hypervariable Region

Site-directed mutagenesis was performed to convert residues within the 204-219 domain of the recombinant B31 OspA to the analogous residues of a European OspA variant, K48. In the region of OspA between residues 204 and 219, which includes the helical domain (amino acids 204-217), there are seven amino acid differences between OspA-B31 and OspA-K48. Three oligonucleotides were generated, each containing nucleotide changes which would incorporate K48 amino acids at their analogous positions in the B31 OspA protein. The oligos used to create the site-directed mutants were: 5'-CTTAATGACTCTGACACTAGTGC-3' (#613, which converts serine at position 204 to threonine, and serine at 206 to threonine (Ser204-Thr, Ser206-Thr)) (SEQ ID NO. 1); 5'-GCTACTAAAAAAACCGGGAAATGGAATTCA-3' (#625, which converts alanine at 214 to glycine, and alanine at 215 to lysine (Ala214-Gly, Ala215-Lys)) (SEQ ID NO. 2); and 5'-GCAGCTTGGGATTCAAAAACATCCACTTTAACA-3' (#640, which converts asparagine at 217 to aspartate, and glycine at 219 to lysine (Asn217-Asp, Gly219-Lys)) (SEQ ID NO. 3).

Site-directed mutagenesis was carried out by performing mutagenesis with pairs of the above oligos. Three site-directed mutants were created, each with two changes: OspA 613 (Ser204-Thr, Ser206-Thr), OspA 625 (Ala214-Gly, Ala215-Lys), and 640 (Asn217-Asp, Gly219-Lys). There were also two proteins with four changes: OspA 613/625 (Ser204-Thr, Ser206-Thr, Ala214-Gly, Ala215-Lys) and OspA 613/640 (Ser204-Thr, Ser206-Thr, Asn217-Asp, Gly219-Lys).

Specificity of Antibody Binding to Epitopes of the Non-mutated Hypervariable Region

Monoclonal antibodies that agglutinate spirochetes, including several which are neutralizing in vitro, recognize epitopes that map to the hypervariable region around Trp216 (Barbour, A. G. et al., Infect. and Immun. 41: 759 (1983); Schubach, W. H. et al., Infect. and Immun. 59: 1911 (1991)). Western Blot analysis demonstrated that chemical cleavage of OspA from the B31 strain at Trp 216 abolishes reactivity of the protein with the agglutinating MAb 105, a monoclonal raised against B31 spirochetes (data not shown). The reagent, n-chlorosuccinimide (NCS), cleaves OspA at the Trp 216, forming a 23.2 kd fragment and a 6.2 kd peptide which is not retained on the Imobilon-P membrane after transfer. The uncleaved material binds MAb 105; however, the 23.2 kd fragment is unreactive. Similar Western blots with a TrpE-OspA fusion protein containing the carboxy-terminal portion of the OspA protein demonstrated that the small 6.2 kd piece also fails to bind MAb 105 (Schubach, W. H. et al., Infect. and Immun. 59: 1911 (1991)).

Monoclonal antibodies H5332 and H3TS (Barbour, A. G. et al., Infect. and Immun. 41: 759 (1983)) have been shown by immunofluorescence to decorate the surface of fixed spirochetes (Wilske, B. et al., World J. Microbiol. 7: 130 (1991)). These monoclonals also inhibit the growth of the organism in culture. Epitope mapping with fusion proteins has confirmed that the epitopes which bind these MAbs are conformationally determined and reside in the carboxy half of the protein. MAb H5332 is cross-reactive among all of the known phylogenetic groups, whereas MAb H3TS and MAb 105 seem to be specific to the B31 strain to which they were raised. Like MAb 105, the reactivities of H5332 and H3TS to OspA are abrogated by fragmentation of the protein at Trp216 (data not shown). MAb 336 was raised to whole spirochetes of the strain PGau. It cross-reacts to OspA from group 1 (the group to which B31 belongs) but not to group 2 (of which K48 is a member). Previous studies using fusion proteins and chemical cleavage have indicated that this antibody recognizes a domain of OspA in the region between residues 217 and 273 (data not shown). All of these MAbs will agglutinate the B31 spirochete.

Western Blot Analysis of Antibody Binding to Mutated Hypervariable Regions

MAbs were used for Western Blot analysis of the site-directed OspA mutants induced in E. coli using the T7 expression system (Dunn, J. J. et al., Protein Expression and Purification 1: 159 (1990)). E. coli cells carrying pET9c plasmids having a site-directed OspA mutant insert were induced at mid-log phase growth with IPTG for four hours at 37.degree. C. Cell lysates were made by boiling an aliquot of the induced cultures in SDS gel loading dye, and this material was then loaded onto a 12% SDS gel (BioRad mini-Protean II), and electrophoresed. The proteins were then transferred to Imobilon-P membranes (Millipore) 70V, 2 hour at 4.degree. C. using the BioRad mini transfer system. Western analysis was carried out as described by Schubach et al. (Infect. Immun. 59: 1911 (1991)).

Western Blot analysis indicated that only the 625 mutant (Ala214-Gly and Ala215-Lys) retained binding to the agglutinating monoclonal H3TS antibody (data not shown). However, the 613/625 mutant which has additional alterations to the amino terminus of Trp216 (Ser204-Thr and Ser206-Thr) did not bind this monoclonal antibody. Both 640 and 613/640 OspAs which have the Asn217-Asp and Gly219-Lys changes on the carboxy-terminal side of Trp216 also failed to bind MAb H3TS. This indicated that the epitope of the B31 OspA which binds MAb H3TS is comprised of amino acid side-chains on both sides of Trp216.

The 613/625 mutant failed to bind MAbs 105 and H5332, while the other mutants retained their ability to bind these MAbs. This is important in light of the data using fusion proteins that indicate that MAb 105 behaves more like MAb H3TS in terms of its serotype specificity and binding to OspA (Wilske, B. et al., Med. Microbiol. Immunol. 181: 191 (1992)). The 613/625 protein has, in addition to the differences at residues Ser204 and Ser206, changes immediately amino-terminal to Trp216 (Ala214-Gly and Ala215-Lys). The abrogation of reactivity of MAbs 105 and H5332 to this protein indicated that the epitopes of OspA which bind these monoclonals are comprised of residues on the amino-terminal side of Trp216.

The two proteins carrying the Asn217-Asp and Gly219-Lys replacements on the carboxy-terminal side of Trp216 (OspAs 640 and 613/640) retained binding to MAbs 105 and H5332; however, they failed to react with MAb 336, a monoclonal which has been mapped with TrpE-OspA fusion proteins and by chemical cleavage to a more carboxy-terminal domain. This result may explain why MAb 336 failed to recognize the K48-type of OspA (Group 2).

It is clear that amino acids Ser204 and Ser206 play an important part in the agglutinating epitopes in the region of the B31 OspA flanking Trp216. Replacement of these two residues altered the epitopes of OspA that bind MAbs 105, H3TS and H5332. The ability of the 640 changes alone to abolish reactivity of MAb 336 indicated that Ser204 and Ser206 are not involved in direct interaction with MAb 336.

The results indicated that the epitopes of OspA which are available to MAbs that agglutinate spirochetes are comprised at least in part by amino acids in the immediate vicinity of Trp216. Since recent circular dichroism analysis indicated that the structures of B31 and K48 OspA differ very little within this domain, it is unlikely that the changes made by mutation have radically altered the overall structure of the OspA protein (France, L. L. et al., Biochem. Biophys. Acta 1120: 59 (1992); and France et al., Biochem. Biophys Acta, submitted (1993)). This hypothesis is supported by the finding that the recombinant, mutant OspAs exhibit the same high solubility and purification properties as the parent B31 protein (data not shown).

In summary, amino acid side-chains at Ser204 and Ser206 are important for many of the agglutinating epitopes. However, a limited set of conservative changes at these sites were not sufficient to abolish binding of all of the agglutinating MAbs. These results suggested that the agglutinating epitopes of OspA are distinct, yet may have some overlap. The results also supported the hypothesis that the surface-exposed epitope around Trp216 which is thought to be important for immune recognition and neutralization is a conformationally-determined and complex domain of OspA.

Example 3

Borrelia Strains and Proteins

Proteins and genes from any strain of Borrelia can be utilized in the current invention. Representative strains are summarized in Table I, above.

A. Genes Encoding Borrelia Proteins

The chimeric peptides of the current invention can comprise peptides derived from any Borrelia proteins. Representative proteins include OspA, OspB, OspC, OspD, p12, p39, p41 (fla), p66, and p93. Nucleic acid sequences encoding several Borrelia proteins are presently available (see Table II, below); alternatively, nucleic acid sequences encoding Borrelia proteins can be isolated and characterized using methods such as those described below.

TABLE-US-00002 TABLE II References for Nucleic Acid Sequences for Several Proteins of Various Borrelia Strains Strain p93 OspA p41 (fla) K48 X69602 (SID 67) X62624 (SID 8) X69610 PGau SID 73 X62387 (SID 10) X69612 (SID 51) DK29 ND X63412 (SID 49) X69608 (SID 53) PKo X69803 (SID 77) X65599 (SID 57) X69613 (SID 131) PTrob X69604 (SID 71) X65598 (SID 135) X69614 Ip3 ND X70365 (SID 56) ND Ip90 ND Kryuchechnikov, ND V. N. et al., J. Microbiol. Epid. Immunobiol. 12: 41-44 (1988) (SID 50) 25015 X70365 (SID 75) Fikrig, E. S. et ND al., J. Immunol. 7: 2256-2260 (1992) (SID 12) B31 Perng, G. C. et al., Bergstrom, S. Gassmann, G. S. Infect. Immun. 59: et al., Mol. et al., Nucl. 2070-74 (1992); Microbiol. Acids Res. 17: Luft, B. J. et al., 3: 479-486 3590 (1989) Infect. Immun. 60: (1989) (SID 6) (SID 127) 4309-4321 (1992) (SID 65) PKa1 ND X69606 (SID 132) X69611 (SID 129) ZS7 ND Jonsson, M. et al., ND Infect. Immun. (60: 1845-1853 (1992) (SID 134) N40 ND Kryuchechnikov, ND V. N. et al. (SID 133) PHei ND X65600 (SID 136) ND ACAI ND Kryuchechnikov, ND V. N. et al. (SID 58) PBo X69601 (SID 69) X65605 (SID 55) X69610 (SID 130) Numbers with an "X" prefix are GenBank data base accession numbers. SID = SEQ ID NO.

B. Isolation of Borrelia Genes

Nucleic acid sequences encoding full length, lipidated proteins from known Borrelia strains were isolated using the polymerase chain reaction (PCR) as described below. In addition, nucleic acid sequences were generated which encoded truncated proteins (proteins in which the lipidation signal has been removed, such as by eliminating the nucleic acid sequence encoding the first 18 amino acids, resulting in non-lipidated proteins). Other proteins were generated which encoded polypeptides of a particular gene (i.e., encoding a segment of the protein which has a different number of amino acids than the protein does in nature). Using similar methods as those described below, primers can be generated from known nucleic acid sequences encoding Borrelia proteins and used to isolate other genes encoding Borrelia proteins. Primers can be designed to amplify all of a gene, as well as to amplify a nucleic acid sequence encoding truncated protein sequences, such as described below for OspC, or nucleic acid sequences encoding a polypeptide derived from a Borrelia protein. Primers can also be designed to incorporate unique restriction enzyme cleavage sites into the amplified nucleic acid sequences. Sequence analysis of the amplified nucleic acid sequences can then be performed using standard techniques.

Cloning and Sequencing of OspA Genes and Relevant Nucleic Acid Sequences

Borrelia OspA sequences were isolated in the following manner: 100 .mu.l reaction mixtures containing 50 mM KCl, 10 mM TRIS-HCl (pH 8,3), 1.5 mM MgCl.sub.2, 200 .mu.M each NTP, 2.5 units of TaqI DNA polymerase (Amplitaq, Perkin-Elmer/Cetus) and 100 pmol each of the 5' and 3' primers (described below) were used. Amplification was performed in a Perkin-Elmer/Cetus thermal cycler as described (Schubach, W. H. et al., Infect. Immun. 59: 1811-1915 (1991)). The amplicon was visualized on an agarose gel by ethidium bromide staining. Twenty nanograms of the chloroform-extracted PCR product were cloned directly into the PC-TA vector (Invitrogen) by following the manufacturer's instructions. Recombinant colonies containing the amplified fragment were selected, the plasmids were prepared, and the nucleic acid sequence of each OspA was determined by the dideoxy chain-termination technique using the Sequenase kit (United States Biochemical). Directed sequencing was performed with M13 primers followed by OspA-specific primers derived from sequences, previously obtained with M13 primers.

Because the 5' and 3' ends of the OspA gene are highly conserved (Fikrig, E. S. et al., J. Immunol. 7: 2256-2260 (1992); Bergstrom, S. et al., Mol. Microbiol. 3: 479-486 (1989); Zumstein, G. et al., Med. Microbiol. Immunol. 181: 57-70 (1992)), the 5' and 3' primers for cloning can be based upon any known OspA sequences. For example, the following primers based upon the OspA nucleic acid sequence from strain B31 were used:

TABLE-US-00003 (SEQ ID NO. 4) 5'-GGAGAATATATTATGAAA-3' (-12 to +6); and (SEQ ID NO. 5) 5'-CTCCTTATTTTAAAGCG-3' (+826 to +809). (Schubach, W. H. et al., Infect. Immun 59: 1811-1915 (1991)).

OspA genes isolated in this manner include those for strains B31, K48, PGau, and 25015; the nucleic acid sequences are depicted in the sequence listing as SEQ ID NO. 6 (OspA-B31), SEQ ID NO. 8 (OspA-K48), SEQ ID NO. 10 (OspA-PGau), and SEQ ID NO. 12 (OspA-25015). An alignment of these and other OspA nucleic acid sequences is shown in FIG. 42. The amino acid sequences of the proteins encoded by these nucleic acid sequences are represented as SEQ ID NO. 7 (OspA-B31), SEQ ID NO. 9 (OspA-K48), SEQ ID NO. 11 (OspA-PGau), and SEQ ID NO. 13 (OspA-25015).

The following primers were used to generate specific nucleic acid sequences of the OspA gene, to be used to generate chimeric nucleic acid sequences (as described in Example 4):

TABLE-US-00004 (SEQ ID NO. 14) 5'-GTCTGCAAAAACCATGACAAG-3' (plus strand primer #369); (SEQ ID NO 15) 5'-GTCATCAACAGAAGAAAAATTC-3' (plus strand primer #357); (SEQ ID NO. 16) 5'-CCGGATCCATATGAAAAAATATTTATTGGG-3' (plus strand primer #607); (SEQ ID NO. 17) 5'-CCGGGATCCATATGGCTAAGCAAAATGTTAGC-3' (plus strand primer #584); (SEQ ID NO. 18) 5'-GCGTTCAAGTACTCCAGA-3' (minus strand primer #200); (SEQ ID NO. 19) 5'-GATATCTAGATCTTATTTTAAAGCGTT-3' (minus strand primer #586); and (SEQ ID NO. 20) 5'-GGATCCGGTGACCTTTTAAAGCGTTTTTAAT-3' (minus strand primer #1169).

Cloning and Sequencing of OspB

Similar methods were also used to isolate OspB genes. One OspB genes isolated is represented as SEQ ID NO. 21 (OspB-B31); its encoded amino acid sequence is SEQ ID NO. 22.

The following primers were used to generate specific nucleic acid sequences of the OspB gene, to be used in generation of chimeric nucleic acid sequences (see Example 4):

TABLE-US-00005 (SEQ ID NO. 23) 5'-GGTACAATTACAGTACAA-3' (plus strand primer #721); (SEQ ID NO. 24) 5'-CCGAGAATCTCATATGGCACAAAAAGGTGCTGAGTCAATTGG-3' (plus strand primer #1105); (SEQ ID NO. 25) 5'-CCGATATCGGATCCTATTTTAAAGCGTTTTTAAGC-3' (minus strand primer #1106); and (SEQ ID NO. 26) 5'-GGATCCGGTGACCTTTTAAAGCGTTTTTAAG-3' (minus strand primer #1170).

Cloning and Sequencing of OspC

Similar methods were also used to isolate OspC genes. The following primers were used to isolate entire OspC genes from Borrelia strains B31, K48, PKo, and PTrob:

TABLE-US-00006 (SEQ ID NO. 27) 5'-GTGCGCGACCATATGAAAAAGAATACATTAAGTGCG-3' (plus strand primer having Ndel site combined with start codon), and (SEQ ID NO. 28) 5'-GTCGGCGGATCCTTAAGGTTTTTTTGGACTTTCTGC-3' (minus strand primer having BamHI site followed by stop codon).

The nucleic acid sequences of the OspC genes were then determined by the dideoxy chain-termination technique using the Sequenase kit (United States Biochemical). OspC genes isolated and sequenced in this manner include those for strains B31, K48, PKo, and Tro; the nucleic acid sequences are depicted in the sequence listing as SEQ ID NO. 29 (OspC-B31), SEQ ID NO. 31 (OspC-K48), SEQ ID NO. 33 (OspC-PKo), and SEQ ID NO. 35 (OspC-Tro). An alignment of these sequences is shown in FIG. 38. The amino acid sequences of the proteins encoded by these nucleic acid sequences are represented as SEQ ID NO. 30 (OspC-B31), SEQ ID NO. 32 (OspC-K48), SEQ ID NO. 34 (OspC-PKo), and SEQ ID NO. 36 (OspC-Tro).

Truncated OspC genes were generated using other primers. These primers were designed to amplify nucleic acid sequences, derived from the OspC gene, that lacked the nucleic acids encoding the signal peptidase sequence of the full-length protein. The primers corresponded to bp 58-75 of the natural protein, with codons for Met-Ala attached ahead.

TABLE-US-00007 For strain B31, the following primer was used: (SEQ ID NO. 37) 5'-GTGCGCGACCATATGGCTAATAATTCAGGGAAAGAT-3'. For strain PKo, (SEQ ID NO. 38) 5'-GTGCGCGACCATATGGCTAGTAATTCAGGGAAAGGT-3' was used. For strains PTrob and K48, (SEQ ID NO. 39) 5'-GTGCGCGACCATATGGCTAATAATTCAGGTGGGGAT-3' was used.

Additional primers were also designed to amplify nucleic acids encoding particular polypeptides, for use in creation of chimeric nucleic acid sequences (see Example 4). These primers included:

TABLE-US-00008 (SEQ ID NO. 40) 5'-CTTGGAAAATTATTTGAA-3' (plus strand primer #520); (SEQ ID NO. 41) 5'-CACGGTCACCCCATGGGAAATAATTCAGGGAAAGG-3' (plus strand primer #58); (SEQ ID NO. 42) 5'-TATAGATGACAGCAACGC-3' (minus strand primer #207); and (SEQ ID NO. 43) 5'-CCGGTGACCCCATGGTACCAGGTTTTTTTGGACTTTCTGC-3' (minus strand primer #636).

Cloning and Sequencing of OspD

Similar methods can be used to isolate OspD genes. An alignment of four OspD nucleic acid sequences (from strains PBo, PGau, DK29, and K48) is shown in FIG. 39.

Cloning and Sequencing of p12

The p12 gene was similarly identified. Primers used to clone the entire p12 gene included: 5'-CCGGATCCATATGGTTAAAAAAATAATATTTATTTC-3' (forward primer # 757) (SEQ ID NO. 44); and 5'-GATATCTAGATCTTTAATTGCTCTGCTCACTCTCTTC-3' (reverse primer #758) (SEQ ID NO. 45).

To amplify a truncated p12 gene (one in which the transcribed protein is non-lipidated, and begins at amino acid 18 of the native sequence), the following primers were used: 5'-CCGGGATCCATATGGCTAGTGCAATTGGTCGTGG-3' (forward primer # 759) (SEQ ID NO. 46); and primer #758 (SEQ ID NO. 45).

Cloning and Sequencing of p41 (fla)

A similar approach was used to clone and sequence genes encoding the p41 (fla) protein. The p41 sequences listed in Table II with GenBank accession numbers were isolated using the following primers from strain B31: 5'-ATGATTATCAATCATAAT-3' (+1 to +18) (SEQ ID NO. 47); and 5'-TCTGAACAATGACAAAAC-3' (+1008 to +991) (SEQ ID NO. 48). The nucleic acid sequences of p41 isolated in this manner are depicted in the sequence listing as SEQ ID NO. 51 (p41-PGau), and SEQ ID NO. 53 (p41-DK29). An alignment of several p41 nucleic acid sequences, including those for strains B31, PKa1, PGau, PBo, DK29, and PKo, is shown in FIG. 41. The amino acid sequences of the proteins encoded by these nucleic acid sequences are represented as SEQ ID NO. 52 (p41-PGau) and SEQ ID NO. 54 (p41-DK29).

Other primers were designed to amplify nucleic acid sequences encoding polypeptides of p41, to be used in chimeric nucleic acid sequences. These primers included:

TABLE-US-00009 (SEQ ID NO. 59) 5'-TTGGATCCGGTCACCCCATGGCTCAATATAACCAATG-3' (minus strand primer #122); (SEQ ID NO. 60) 5'-TTGGATCCGGTCACCCCATGGCTTCTCAAAATGTAAG-3' (plus strand primer #140); (SEQ ID NO. 61) 5'-TTGGATCCGGTGACCAACTCCGCCTTGAGAAGG-3' (minus strand primer #234); and (SEQ ID NO. 62) 5'-TTGGATCCGGTGACCTATTTGAGCATAAGATGC-3' (minus strand primer #141).

Cloning and Sequencing of p93

The same approach was also used to clone and sequence p93 proteins. Genes encoding p93, as listed in Table II with GenBank accession numbers, were isolated by this method with the following primers from strain B31:

TABLE-US-00010 (SEQ ID NO. 63) 5'-GGTGAATTTAGTTGGTAAGG-3' (-54 to -35); and (SEQ ID NO. 64) 5'-CACCAGTTTCTTTAAGCTGCTCCTGC-3' (+1117 to +1092).

The nucleic acid sequences of p93 isolated in this manner are depicted in the sequence listing as SEQ ID NO. 65 (p93-B31), SEQ ID NO. 67 (p93-K48) SEQ ID NO. 69 (p93-PBo), SEQ ID NO. 71 (p93-PTrob), SEQ ID NO. 73 (p93-PGau), SEQ ID NO. 77 (p93-25015), and SEQ ID NO. 75 (p93-PKo). The amino acid sequences of the proteins encoded by these nucleic acid sequences are represented as SEQ ID NO. 66 (p93-B31), SEQ ID NO. 68 (p93-K48) SEQ ID NO. 70 (p93-PBo), SEQ ID NO. 72 (p93-PTrob), SEQ ID NO. 74 (p93-PGau), SEQ ID NO. 78 (p93-25015), and SEQ ID NO. 76 (p93-PKo).

Other primers were used to amplify nucleic acid sequences encoding polypeptides of p93 to be used in generating chimeric nucleic acid sequences. These primers included:

TABLE-US-00011 (SEQ ID NO. 79) 5'-CCGGTCACCCCATGGCTGCTTTAAAGTCTTTA-3' (plus strand primer #475); (SEQ ID NO. 80) 5'-CCGGTCACCCCATGAATCTTGATAAAGCTCAG-3' (plus strand primer #900); (SEQ ID NO. 81) 5'-CCGGTCACCCCATGGATGAAAAGCTTTTAAAAAGT-3' (plus strand primer #1168); (SEQ ID NO. 82) 5'-CCGGTCACCCCCATGGTTGAGAAATTAGATAAG-3' (plus strand primer #1423); and (SEQ ID NO. 83) 5'-TTGGATCCGGTGACCCTTAACTTTTTTTAAAG-3' (minus strand primer #2100).

C. Expression of Proteins from Borrelia Genes

The nucleic acid sequences described above can be incorporated into expression plasmids, using standard techniques, and transfected into compatible host cells in order to express the proteins encoded by the nucleic acid sequences. As an example, the expression of the p12 gene and the isolation of p12 protein is set forth.

Amplification of the p12 nucleic acid sequence was conducted with primers that included a NdeI restriction site into the nucleic acid sequence. The PCR product was extracted with phenol/chloroform and precipitated with ethanol. The precipitated product was digested and ligated into an expression plasmid as follows: 15 .mu.l (approximately 1 .mu.g) of PCR DNA was combined with 2 .mu.l 10.times. restriction buffer for NdeI (Gibco/BRL), 1 .mu.l NdeI (Gibco/BRL), and 2 .mu.l distilled water, and incubated overnight at 37.degree. C. This mixture was subsequently combined with 3 .mu.l 10.times. buffer (buffer 3, New England BioLabs), 1 .mu.l BamHI (NEB), and 6 .mu.l distilled water, and incubated at 37.degree. for two hours. The resultant material was purified by preparative gel electrophoresis using low melting point agarose, and the band was visualized under long wave ultraviolet light and excised from the gel. The gel slice was treated with Gelase using conditions recommended by the manufacturer (Epicentre Technologies). The resulting DNA pellet was resuspended in 25-50 .mu.l of 10 mM TRIS-CL (pH 8.0) and 1 mM EDTA (TE). An aliquot of this material was ligated into the pET9c expression vector (Dunn, J. J. et al., Protein Expression and Purification 1: 159 (1990)).

To ligate the material into the pET9c expression vector, 20-50 ng of p12 nucleic acid sequences cut and purified as described above was combined with 5 .mu.l 10 One-Phor-All (OPA) buffer (Pharmacia), 30-60 ng pET9c cut with NdeI and BamHI, 2.5 .mu.l 20 mM ATP, 2 .mu.l T4 DNA ligase (Pharmacia) diluted 1:5 in 1.times. OPA buffer, and sufficient distilled water to bring the final volume to 50 .mu.l. The mixture was incubated at 12.degree. C. overnight.

The resultant ligations were transformed into competent DH5-alpha cells and plated on nutrient agar plates containing 50 .mu.g/ml kanamycin and incubated overnight at 37.degree. C. DH5-alpha is used as a "storage strain" for T7 expression clones, because it is RecA deficient, so that recombination and concatenation are not problematic, and because it lacks the T7 RNA polymerase gene necessary to express the cloned gene. The use of this strain allows for cloning of potentially toxic gene products while minimizing the chance of deletion and/or rearrangement of the desired genes. Other cell lines having similar properties may also be used.

Kanamycin resistant colonies were single-colony purified on nutrient agar plates supplemented with kanamycin at 50 .mu.g/ml. A colony from each isolate was inoculated into 3-5 ml of liquid medium containing 50 .mu.g/ml kanamycin, and incubated at 37.degree. C. without agitation. Plasmid DNA was obtained from 1 ml of each isolate using a hot alkaline lysis procedure (Mantiatis, T. et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. (1982)).

Plasmid DNA was digested with EcoRI and BglII in the following manner: 15 .mu.l plasmid DNA was combined with 2 .mu.l 10.times. buffer 3 (NEB), 1 .mu.l EcoRI (NEB), 1 .mu.l BglII (NEB) and 1 .mu.l distilled water, and incubated for two hours at 37.degree. C. The entire reaction mixture was electrophoresed on an analytical agarose gel. Plasmids carrying the p12 insert were identified by the presence of a band corresponding to 925 base-pairs (full length p12) or 875 base-pairs (nonlipidated p12). One or two plasmid DNAs from the full length and nonlipidated p12 clones in pET9c were used to transform BL21 DE3 pLysS to kanamycin resistance as described by Studier et al. (Methods in Enzymology, Goeddel, D. (Ed.), Academic Press, 185: 60-89 (1990)). One or two transformants of the full length and nonlipidated clones were single-colony purified on nutrient plates containing 25 .mu.g/ml chloramphenicol (to maintain pLysS) and 50 .mu.g/ml kanamycin at 37.degree. C. One colony of each isolate was inoculated into liquid medium supplemented with chloramphenicol and kanamycin and incubated overnight at 37.degree. C. The overnight culture was subcultured the following morning into 500 ml of liquid broth with chloramphenicol (25 .mu.g/ml) and kanamycin (50 .mu.g/ml) and grown with aeration at 37.degree. C. in an orbital air-shaker until the absorbance at 600 nm reached 0.4-0.7. Isopropyl-thio-galactoside (IPTG) was added to a final concentration of 0.5 mM, for induction, and the culture was incubated for 3-4 hours at 37.degree. C. as before. The induced cells were pelleted by centrifugation and resuspended in 25 ml of 20 mM NaPO.sub.4 (pH 7.7). A small aliquot was removed for analysis by gel electrophoresis. Expressing clones produced proteins which migrated at the 12 kDa position.

A crude cell lysate was prepared from the culture as described for recombinant OspA by Dunn, J. J. et al., (Protein Expression and Purification 1: 159 (1990)). The crude lysate was first passed over a Q-sepharose column (Pharmacia) which had been pre-equilibrated in Buffer A: 10 mM NaPO.sub.4 (pH 7.7), 10 mM NaCl, 0.5 mM PMSF. The column was washed with 10 mM NaPO.sub.4, 50 mM NaCl and 0.5 mM PMSF and then p12 was eluted in 10 mM NaPO.sub.4, 0.5 mM PMSF with a NaCl gradient from 50-400 mM. p12 eluted approximately halfway through the gradient between 100 and 200 mM NaCl. The peak fractions were pooled and dialyzed against 10 mM NaPO.sub.4 (pH 7.7), 10 mM NaCl, 0.5 mM PMSF. The protein was then concentrated and applied to a Sephadex G50 gel filtration column of approximately 50 ml bed volume (Pharmacia), in 10 mM NaPO.sub.4, 200 mM NaCl, 0.5 mM PMSF. p12 would typically elute shortly after the excluded volume marker. Peak fractions were determined by running small aliquots of all fractions on an SDS gel. The p12 peak was pooled and stored in small aliquots at -20.degree. C.

Example 4

Generation of Chimeric Nucleic Acid Sequences and Chimeric Proteins

A. General Protocol for Creation of Chimeric Nucleic Acid Sequences

The megaprimer method of site directed mutagenesis and its modification were used to generate chimeric nucleic acid sequences (Sarkar and Sommer, Biotechniques 8(4): 404-407 (1990); Aiyar, A. and J. Leis, Biotechniques 14(3): 366-369 (1993)). A 5' primer for the first genomic template and a 3' fusion oligo are used to amplify the desired region. The fusion primer consists of a 3' end of the first template (DNA that encodes the amino-proximal polypeptide of the fusion protein), coupled to a 5' end of the second template (DNA that encodes the carboxy-proximal polypeptide of the fusion protein).

The PCR amplifications are performed using Taq DNA polymerase, 10.times.PCR buffer, and MgCl.sub.2 (Promega Corp., Madison, Wis.), and Ultrapure dNTPs (Pharmacia, Piscataway, N.J.). One .mu.g of genomic template 1, 5 .mu.l of 10 .mu.M 5' oligo and 5 .mu.l of 10 .mu.M fusion oligo are combined with the following reagents at indicated final concentrations: 10.times. Buffer-Mg FREE (1.times.), MgCl.sub.2 (2 mM), dNTP mix (200 .mu.M each dNTP), Taq DNA polymerase (2.5 units), water to bring final volume to 100 .mu.l. A Thermal Cycler (Perkin Elmer Cetus, Norwalk, Conn.) is used to amplify under the following conditions: 35 cycles at 95.degree. C. for one minute, 55.degree. C. for two minutes, and 72.degree. for three minutes. This procedure results in a "megaprimer".

The resulting megaprimer is run on a 1.times.TAE, 4% low-melt agarose gel. The megaprimer band is cut from the gel and purified using the Promega Magic PCR Preps DNA purification system. Purified megaprimer is then used in a second PCR step. One .mu.g of genomic template 2, approximately 0.5 .mu.g of the megaprimer, and 5 .mu.l of 10 .mu.M 3' oligo are added to a cocktail of 10.times. buffer, MgCl.sub.2, dNTPs and Taq at the same final concentrations as noted above, and brought to 100 .mu.l with water. PCR conditions are the same as above. The fusion product resulting from this amplification is also purified using the Promega Magic PCR Preps DNA purification system.

The fusion product is then ligated into TA vector and transformed into E. coli using the Invitrogen (San Diego, Calif.) TA Cloning Kit. Approximately 50 ng of PCR fusion product is ligated to 50 ng of pCRII vector with 1.times. Ligation Buffer, 4 units of T4 ligase, and brought to 10 .mu.l with water. This ligated product mixture is incubated at 12.degree. C. overnight (approximately 14 hours). Two .mu.l of the ligation product mixture is added to 50 .mu.l competent INC F' cells and 2 .mu.l beta mercaptoethanol. The cells are then incubated for 30 minutes, followed by heat shock treatment at 42.degree. C. for 60 seconds, and an ice quenching for two minutes. 450 .mu.l of warmed SOC media is then added to the cells, resulting in a transformed cell culture which is incubated at 37.degree. C. for one hour with slight shaking. 50 .mu.l of the transformed cell culture is plated on LB+50 .mu.g/.mu.l ampicillin plates and incubated overnight at 37.degree. C. Single white colonies are picked and added to individual overnight cultures containing 3 ml LB with ampicillin (50 .mu.g/.mu.l).

The individual overnight cultures are prepared using Promega's Magic Miniprep DNA purification system. A small amount of the resulting DNA is cut using a restriction digest as a check. DNA sequencing is then performed to check the sequence of the fusion nucleic acid sequence, using the United States Biochemical (Cleveland, Ohio) Sequenase Version 2.0 DNA sequencing kit. Three to five .mu.g of plasmid DNA is used per reaction. 2 .mu.l 2M NaOH/2 mM EDTA are added to the DNA, and the volume is brought to 20 .mu.l with water. The mixture is then incubated at room temperature for five minutes. 7 .mu.l water, 3 .mu.l 3M NaAc, 75 .mu.l EtOH are added. The resultant mixture is mixed by vortex and incubated for ten minutes at -70.degree. C., and then subjected to microcentrifugation. After microcentrifugation for ten minutes, the supernatant is aspirated off, and the pellet is dried in the speed vac for 30 second. 6 .mu.l water, 2 .mu.l annealing buffer, and 2 .mu.l of 10 .mu.M of the appropriate oligo is then added. This mixture is incubated for 10 minutes at 37.degree. C. and then allowed to stand at room temperature for 10 minutes. Subsequently, 5.5 .mu.l of label cocktail (described above) is added to each sample of the mixture, which are incubated at room temperature for an additional five minutes. 3.5 .mu.l labeled DNA is then added to each sample which is then incubated for five minutes at 37.degree. C. 4 .mu.l stop solution is added to each well. The DNA is denatured at 95.degree. for two minutes, and then placed on ice.

Clones with the desired fusion nucleic acid sequences are then reckoned in frame in the pET expression system in the lipidated (full length) and non-lipidated (truncated, i.e., without first 17 amino acids) forms. The product is amplified using restriction sites contained in the PCR primers. The vector and product are cut with the same enzymes and ligated together with T4 ligase. The resultant plasmid is transformed into competent E. coli using standard transformation techniques. Colonies are screened as described earlier and positive clones are transformed into expression cells, such as E. coli BL21, for protein expression with IPTG for induction. The expressed protein in its bacterial culture lysate form and/or purified form is then injected in mice for antibody production. The mice are bled, and the sera collected for agglutination, in vitro growth inhibition, and complement-dependent and -independent lysis tests.

B. Specific Chimeric Nucleic Acid Sequences

Various chimeric nucleic acid sequences were generated. The nucleic acid sequences are described as encoding polypeptides from Borrelia proteins. The chimeric nucleic acid sequences are produced such that the nucleic acid sequence encoding one polypeptide is in the same reading frame as the nucleic acid sequence encoding the next polypeptide in the chimeric protein sequence encoded by the chimeric nucleic acid sequence. The proteins are listed sequentially (in order of presence of the encoding sequence) in the description of the chimeric nucleic acid sequence. For example, if a chimeric nucleic acid sequence consists of bp 1-650 from OspA-1 and bp 651-820 from OspA-2 were sequenced, the sequence of the chimer would include the first 650 base pairs from OspA-1 followed immediately by base pairs 651-820 of OspA-2.

OspA-K48/OspA-PGau

A chimer of OspA from strain K48 (OspA-K48) and OspA from strain PGau (OspA-PGau) was generated using the method described above. This chimeric nucleic acid sequence included bp 1-654 from OspA-K48, followed by bp 655-820 from OspA-PGau. Primers used included: the amino-terminal sequence of OspA primer #607 (SEQ ID NO. 16); the fusion primer, 5'-AAAGTAGAAGTTTTTGAATCCCATTTTCCAGTTTTTTT-3' (minus strand primer #668-654) (SEQ ID NO. 84); the carboxy-terminal sequence of OspA primer #586 (SEQ ID NO. 19); and the sequence primers #369 (SEQ ID NO. 14) and #357 (SEQ ID NO. 15). The chimeric nucleic acid sequence is presented as SEQ ID NO. 85; the chimeric protein encoded by this chimeric nucleic acid sequence is presented as SEQ ID NO. 86.

OspA-B31/OspA-PGau

A chimer of OspA from strain B31 (OspA-B31) and OspA from strain PGau (OspA-PGau) was generated using the method described above. This chimeric nucleic acid sequence included bp 1-651 from OspA-B31, followed by bp 652-820 from OspA-PGau. Primers used included: the fusion primer, 5'-AAAGTAGAAGTTTTTGAATTCCAAGCTGCAGTTTT-3' (minus strand primer #668-651) (SEQ ID NO. 87); and the sequence primer, #369 (SEQ ID NO. 14). The chimeric nucleic acid sequence is presented as SEQ ID NO. 88; the chimeric protein encoded by this chimeric nucleic acid sequence is presented as SEQ ID NO. 89. OspA-B31/OspA-K48

A chimer of OspA from strain B31 (OspA-B31) and OspA from strain K48 (OspA-K48) was generated using the method described above. This chimeric nucleic acid sequence included bp 1-651 from OspA-B31, followed by bp 652-820 from OspA-K48. Primers used included: the fusion primer,

5'-AAAGTGGAAGTTTTTGAATTCCAAGCTGCAGTTTTTTT-3' (minus strand primer #671-651) (SEQ ID NO. 90); and the sequence primer, #369 (SEQ ID NO. 14). The chimeric nucleic acid sequence is presented as SEQ ID NO. 91; the chimeric protein encoded by this chimeric nucleic acid sequence is presented as SEQ ID NO. 92. OspA-B31/OspA-25015

A chimer of OspA from strain B31 (OspA-B31) and OspA from strain 25015 (OspA-25015) was generated using the method described above. This chimeric nucleic acid sequence included bp 1-651 from OspA-B31, followed by bp 652-820 from OspA-25015. Primers used included: the fusion primer, 5'-TAAAGTTGAAGTGCCTGCATTCCAAGCTGCAGTTT-3' (SEQ ID NO. 93). The chimeric nucleic acid sequence is presented as SEQ ID NO. 94; the chimeric protein encoded by this chimeric nucleic acid sequence is presented as SEQ ID NO. 95.

OspA-K48/OspA-B31/OspA-K48

A chimer of OspA from strain B31 (OspA-B31) and OspA from strain K48 (OspA-K48) was generated using the method described above. This chimeric nucleic acid sequence included bp 1-570 from OspA-K48, followed by bp 570-651 from OspA-B31, followed by bp 650-820 from OspA-K48. Primers used included: the fusion primer, 5'-CCCCAGATTTTGAAATCTTGCTTAAAACAAC-3' (SEQ ID NO. 96); and the sequence primer, #357 (SEQ ID NO. 15). The chimeric nucleic acid sequence is presented as SEQ ID NO. 97; the chimeric protein encoded by this chimeric nucleic acid sequence is presented as SEQ ID NO. 98.

OspA-B31/OspA-K48/OspA-B31/OspA-K48

A chimer of OspA from strain B31 (OspA-B31) and OspA from strain K48 (OspA-K48) was generated using the method described above. This chimeric nucleic acid sequence included bp 1-420 from OspA-B31, followed by 420-570 from OspA-K48, followed by bp 570-650 from OspA-B31, followed by bp 651-820 from OspA-K48. Primers used included: the fusion primer, 5'CAAGTCTGGTTCCAATTTGCTCTTGTTATTAT-3' (minus strand primer #436-420) (SEQ ID NO. 99); and the sequence primer, #357 (SEQ ID NO. 15). The chimeric nucleic acid sequence is presented as SEQ ID NO. 100; the chimeric protein encoded by this chimeric nucleic acid sequence is presented as SEQ ID NO. 101.

OspA-B31/OspB-B31

A chimer of OspA and OspB from strain B31 (OspA-B31, OspB-B31) was generated using the method described above. The chimeric nucleic acid sequence included bp 1-651 from OspA-B31, followed by bp 652-820 from OspB-B31. Primers used included: the fusion primer, 5'-GTTAAAGTGCTAGTACTGTCATTCCAAGCTGCAGTTTTTTT-3' (minus strand primer #740-651) (SEQ ID NO. 102); the carboxy-terminal sequence of OspB primer #1106 (SEQ ID NO. 25); and the sequence primer #357 (SEQ ID NO. 15). The chimeric nucleic acid sequence is presented as SEQ ID NO. 103; the chimeric protein encoded by this chimeric nucleic acid sequence is presented as SEQ ID NO. 104.

OspA-B31/OspB-B31/OspC-B31

A chimer of OspA, OspB and OspC from strain B31 (OspA-B31, OspB-B31, and OspC-B31) was generated using the method described above. The chimeric nucleic acid sequence included bp 1-650 from OspA-B31, followed by bp 652-820 from OspB-B31, followed by bp 74-630 of OspC-B31. Primers used included: the fusion primer, 5'-TGCAGATGTAATCCCATCCGCCATTTTTAAAGCGTTTTT-3' (SEQ ID NO. 105); and the carboxy-terminal sequence of OspC primer (SEQ ID NO. 28). The chimeric nucleic acid sequence is presented as SEQ ID NO. 106; the chimeric protein encoded by this chimeric nucleic acid sequence is presented as SEQ ID NO. 107.

OspC-B31/OspA-B31/OspB-B31

A chimer of OspA, OspB and OspC from strain B31 (OspA-B31, OspB-B31, and OspC-B31) was generated using the method described above. The chimeric nucleic acid sequence included bp 1-630 from OspC-B31, followed by bp 52-650 from OspA-B31, followed by bp 650-820 of OspB-B31. Primers used included: the amino-terminal sequence of OspC primer having SEQ ID NO. 27; the fusion primer, 5'-GCTGCTAACATTTTGCTTAGGTTTTTTTGGACTTTC-3' (minus strand primer #69-630) (SEQ ID NO. 108); and the sequence primers #520 (SEQ ID NO. 40) and #200 (SEQ ID NO. 18). The chimeric nucleic acid sequence is presented as SEQ ID NO. 109; the chimeric protein encoded by this chimeric nucleic acid sequence is presented as SEQ ID NO. 110.

Additional Chimeric Nucleic Acid Sequences

Using the methods described above, other chimeric nucleic acid sequences were produced. These chimeric nucleic acid sequences, and the proteins encoded, are summarized in Table III.

TABLE-US-00012 TABLE III Chimeric Nucleic acid Sequences and the Encoded Proteins SEQ ID NO. SEQ ID NO. Chimers Generated (base pairs) (nt) (protein) OspA (52-882)/p93 (1168-2100) 111 112 OspB (45-891)/p41 (122-234) 113 114 OspB (45-891)/p41 (122-295) 115 116 OspB (45-891)/p41 (140-234) 117 118 OspB (45-891)/p41 (140-295) 119 120 OspB (45-891)/p41 (122-234)/ 121 122 OspC (58-633) OspA-Tro/OspA-Bo 137 138 OspA-PGau/OspA-Bo 139 140 OspA-B31/OspA-PGau/OspA-B31/ 143 144 OspA-K48 OspA-PGau/OspA-B31/OspA-K48 141 142

C. Purification of Proteins Generated by Chimeric Nucleic Acid Sequences

The chimeric nucleic acid sequences described above, as well as chimeric nucleic acid sequences produced by the methods described above, are used to produce chimeric proteins encoded by the nucleic acid sequences. Standard methods, such as those described above in Example 3, concerning the expression of proteins from Borrelia genes, can be used to express the proteins in a compatible host organism. The chimeric proteins can then be isolated and purified using standard techniques.

If the chimeric protein is soluble, it can be purified on a Sepharose column. Insoluble proteins can be solubilized in guanidine and purified on a Ni.sup.2+ column; alternatively, they can be solubilized in 10 mM NaPO.sub.4 with 0.1-1% TRIXON X 114, and subsequently purified over an S column (Pharmacia). Lipidated proteins were generally purified by the latter method. Solubility was determined by separating both soluble and insoluble fractions of cell lysate on a 12% PAGE gel, and checking for the localization of the protein by Coomasie staining, or by Western blotting with monoclonal antibodies directed to an antigenic polypeptide of the chimeric protein.

Example 5

Generation of OspC/OspA Chimeric Nucleic Acids and Chimeric Proteins

A. General Protocol for Creation of Chimeric Nucleic Acid Sequences

A large number of chimeric nucleic acid sequences encoding proteins comprising at least a first and a second polypeptide from Borrelia burgdorferi were generated. These chimeric nucleic acid sequences were produced such that the encoded chimeric protein comprised a Borrelia burgdorferi OspC polypeptide upstream of (or N-terminal to) a Borrelia burgdorferi OspA polypeptide. The chimeric nucleic acid sequences were also produced such that the nucleic acid encoding one polypeptide was in the same reading frame as the nucleic acid sequence encoding the next polypeptide in the chimeric protein.

The general cloning strategy used to construct the chimeric nucleic acid sequences was as follows. The desired fragment of OspC was amplified using a 5' primer containing a restriction site suitable for cloning the resultant product into a vector of interest and a 3' primer containing a restriction site suitable for ligating the OspC fragment to the OspA fragment. The OspC product is cloned into a suitable vector. For the OspA poriton of the chimeric nucleic acid, the desired OspA fragment was amplified using a 5' primer containing a restriction site for ligating the resultant OspA fragment to the OspC fragment and a 3' primer containing a restriction site suitable for cloning the resultant OspA product into the vector with the OspC product. The use of a restriction site to allow ligation of the OspC and OspA fragment results in the insertion of 0 to about 3 amino acids between the OspC and OspA fragments.

A specific example of such a construction follows. It is understood that other suitable restriction sites could be used with no more than routine experimentation. The resultant OspC/OspA chimers could have, therefore, the addition of 0 to about 3 amino acids between the OspC and OspA fragments, depending on the restriction site used.

For the OspC portions of the chimeric nucleic acids, desired fragments of OspC genes from various strains or genospecies were PCR amplified using a 5' primer containing an NdeI site and a 3' primer containing a NcoI and a BamHI site. The amplified OspC product was then cloned into the NdeI and BamHI sites of the T7 promoter driven expression vector, pET9c. For the OspA portion of the chimeric nucleic acid, desired fragments of OspA genes a strain of interest or genospecies of interest were PCR amplified using a 5' primer containing an NcoI site and a 3' primer containing a BamHI site. This OspA portion could then be directly cloned into the NcoI and BamHI sites of the pET9c vector containing the desired OspC sequence, thereby producing the desired OspC-OspA construct. By including the sequence for the NcoI restriction site in the primers, a nine nucleotide linker sequence encoding the amino acids Ser-Met-Ala was produced at the junction between the N-terminal OspC sequence and the C-terminal OspA sequence. The use of the NcoI restriction enzyme (CCATGG) in this cloning strategy was a suitable choice as Borrelia DNA is an AT-rich and therefore possesses only a few NcoI sites in its genome.

As an example, OspC-OspA chimeric nucleic acids which contain unlipidated OspC B31 were generated using the following primers:

TABLE-US-00013 (5'OspC-NdeI): (SEQ ID NO: 181) 5'-GT CAT ATG GCT TGT AAT AAT TCA GGG AAA GA-3'; and (3'OspC-Ncol): (SEQ ID NO: 182) 5'-T TTC CAT GGA AGG TTT TTT TGG ACT TTC TG-3'.

For OspC-OspA chimeric nucleic acids which contain unlipidated OspA B31, the following primers were used:

TABLE-US-00014 (5'OspA-NcoI:) (SEQ ID NO: 183) 5'-TT TCC ATG GCC AAG CAA AAT GTT AGC AGC C-3'; and (3'OspA-BamHI): (SEQ ID NO: 184) 5'-TAA GGA TCC TTA TTT TAA AGC GTT TTT-3'.

Lipidated versions of the OspC/A chimeras can be constructed by cloning an expression vector that contains a leader sequence containing a lipidation site, such that the leader sequence is linked upstream of the OspC portion of the chimera and in frame with the chimera. The leader sequence comprising a lipidation signal can be, for example, from a gene encoding the OspA, B or C polypeptides.

Chimeric nucleic acid sequences, and the proteins that they encode, which were produced are summarized in Table IV. Other additional chimeric nucleic acid sequences, and encoded proteins, are also depicted in Table IV. In further embodiments, chimeric OspC/OspA proteins are constructed wherein a first segment of OspA is from B31 and comprises base pairs from about 88 to about 450, and a second segment of OspA comprises base pairs from about 451 to about 537 of PKo. These chimeras can also comprise additional OspA segments such as the last two segments of SEQ ID NOs 167 or 165 or the last segment of SEQ ID NO: 163.

TABLE-US-00015 TABLE IV Chimeric OspC/OspA Nucleic Acid Sequences and Encoded Proteins SEQ ID NO. SEQ ID NO. .sup.1Chimers Generated (nt) (protein) FIG. NO. OspC-B31(bp55-633)/OspA-B31(bp52-822) 145 55 OspC-B31(aa19-211)/OspA-B31(aa18-273) 146 55 OspC-B31(bp55-624)/OspA-B31(bp52-822) 147 56 OspC-B31(aa19-208)/OspA-B31(aa18-273) 148 56 OspC-C2(bp55-612)/OspA-B31(bp52-822) 149 57 OspC-C2(aa19-204)/OspA-B31(aa18-273) 150 57 OspC-B31(bp55-633)/OspA-B31(bp52-651)/OspA-K48(bp652-820) 151 58 OspC-B31(aa19-211)/OspA-B31(aa18-216)/OspA-K48(aa217-273) 152 58 OspC-C2(bp55-612)/OspA-B31(bp52-651)/OspA-K48(bp652-820) 153 59 OspC-C2(aa19-204)/OspA-B31(aa18-216)/OspA-K48(aa217-273) 154 59 OspC-B31(bp55-633)/OspA-B31(bp52-651)/OspA-PKo(bp652-820) 155 60 OspC-B31(aa19-211)/OspA-B31(aa18-216)/OspA-PKo(aa217-273) 156 60 OspC-C2(bp55-612)/OspA-B31(52-651)/OspA-PKo(bp652-820) 157 61 OspC-C2(aa19-204)/OspA-B31(aa18-216)/OspA-PKo(aa217-273) 158 61 OspC-B31(bp55-633)/OspA-K48(bp52-654)/OspA-Tro(bp655-819) 159 62 OspC-B31(aa19-211)/OspA-K48(aa18-217)/OspA-Tro(aa218-273) 160 62 OspC-C2(bp55-612/OspA-K48(bp52-654)/OspA-Tro(bp655-819) 161 63 OspC-C2(aa19-204)/OspA-K48(aa18-217)/OspA-Tro(aa218-273) 162 63 OspC-C12(bp55-612)/OspA-B31(bp88-492)/OspA-PKo(bp493-537)/OspA-B31(bp538-8- 22) 163 64 OspC-C12(aa19-204)/OspA-B31(aa30-164)/OspA-PKo(aa165-179)/OspA-B31(aa180-2- 73) 164 64 OspC-PKo(bp55-639)/OspA-B31(bp88-492)/OspA-PKo(bp493-537)/ 165 65 OspA-B31(bp538-651)/OspA-K48(bp652-825) OspC-PKo(aa19-213)/OspA-B31(aa30-164)/OspA-PKo(aa165-179)/ 166 65 OspA-B31(aa180-216)/OspA-K48(aa217-274) OspC-Tro(bp55-624)/OspA-B31(bp88-492)/OspA-PKo(bp493-537)/ 167 66 OspA-B31(bp538-651)/OspA-PKo(bp652-822) OspC-Tro(aa19-208)/OspA-B31(aa30-164)/OspA-PKo(aa165-179)/ 168 66 OspA-B31(aa180-216)/OspA-PKo(aa217-273) OspC-B31(bp55-633)/OspA-B31(bp394-820) 169 67 OspC-B31(aa19-211)/OspA-B31(aa132-273) 170 67 OspC-B31(bp55-631)/OspA-B31(bp394-651)/OspA-K48(652-820) 171 68 OspC-B31(aa19-211)/OspA-B31(aa132-216)/OspA-K48(217-273) 172 68 OspC-B31(bp55-633)/OspA-B31(bp394-651)/OspA-PKo(652-820) 173 69 OspC-B31(aa19-211)/OspA-B31(aa132-216)/OspA-PKo(217-273) 174 69 OspC-B31(bp55-633)/OspA-K48(bp394-654)/OspA-Tro(655-819) 175 70 OspC-B31(aa19-211)/OspA-K48(aa132-217)/OspA-Tro(218-273) 176 70 OspC-B31(bp55-633)/OspA-B31(bp88-492)/OspA-PKo(bp493-537)/ 177 71 OspA-B31(bp541-651)/OspA-PKo(bp652-822) OspC-B31(aa19-211)/OspA-B31(aa30-164)/OspA-PKo(aa165-179, 178 71 aa164(K > G))/OspA-B31(aa180-216)(aa190(N-del))/OspA-PKo(aa217-273) OspC-C2(bp55-612)/OspA-B31(bp88-492)/OspA-PKo(bp493-537)/ 179 72 OspA-B31(bp541-651)/OspA-PKo(bp652-822) OspC-C2(aa19-204)/OspA-B31(aa30-164)/OspA-PKo(aa165-179, 180 72 aa164(K > G))/OspA-B31(aa180-216)(aa190(N-del))/OspA-PKo(aa217-273) .sup.1Chimers Generated are listed as follows: Nucleotide or polypeptide fragment -strain (sequence in base pairs for the top listing and amino acids for the bottom listing) Separate nucleotide or polypeptide fragments in the chimer are separated by a /

B. Protein Expression

As described in the previous two examples, it is possible to express and purify Borrelia proteins such as OspA, OspC and chimeric OspC/OspA polypeptides. This is accomplished by incorporating the desired nucleic acid sequence, which encodes the protein of choice, into an expression plasmid, using standard techniques. This expression plasmid can then be transfected into a compatible host cell in order to express the desired protein.

The purified OspA, OspC or OspC/OspA chimeric proteins, that were used to immunize mice and in the ELISA tests described below, were generated and purified by cloning either OspA, OspC or OspC/OspA chimeric nucleic acid sequences, in frame, into the pET expression plasmid. The expression plasmid was then transfected into the compatible expression cell line Escherichia coli strain BL21 (DE3)/(pLysS) or strain B834 (DE3). The BL21 or B834 cells were then grown in 10 ml LB media (5 g/l NaCl, 10 g/l tryptone, 5 g/l yeast extract, 25 mg/l chloramphenicol and 50 mg/l ampicillin) at 37.degree. C., with shaking. When the optical density at 600% reached 0.3-0.4 units, recombinant protein expression was induced by adding IPTG (isopropyl B-D-thiogalactopyranoside) to a final concentration of 0.5 mM and the cells were grown for an additional three hours. The cultures were harvested by centrifugation at 3800.times.g for five minutes. The cells were resuspended in 20 mM NaPO.sub.4, pH7.7 and stored at -20.degree. C. overnight. Once thawed, the crude extracts were incubated with DNase (2 .mu.g/ml) in the presence of 2.5 mM MgCl.sub.2 at room temperature for thirty minutes and then spun at 14,000 rpm (Eppendorf 5417C) for five minutes.

To purify the OspC proteins described below, the crude extracts from the OspC-expressing cells were loaded onto an anion exchange column (Q Sepharose Fast Flow, 2.2.times.10 cm, Pharmacia) which had been pre-equilibrated with 20 mM Tris-Cl, pH 9.3. The column was washed in the same buffer (20 mM Tris-Cl, pH 9.3) which eluted the OspC protein. The wash fractions that contained OspC were concentrated using Amicon 10K and then were dialyzed with a solution containing 20 mM NaPO.sub.4 pH 8.0 and 250 mM NaCl. The partially purified OspC was then passed over a Ni.sup.2+ metal affinity column (Chelating Sepharose Fast Flow 2.2.times.10 cm, Pharmacia) equilibrated with 20 mM NaPO.sub.4 pH 8.0 and 250 mM NaCl. The column was washed using a decreasing pH gradient of 20 mM sodium acetic acid and 250 mM NaCl and the bound OspC eluted around pH 5.7. The OspC fractions were then concentrated by ultrafiltration and stored at -70.degree. C.

For purification of OspA proteins, the same procedure was followed, except that the dialysis step, after the Amicon 10K cutoff, was done in 20 nM NaPO.sub.4, pH 6.0. The partially purified OspA was then applied to a cation exchange column (S Sepharose Fast Flow 2.2.times.10 cm, Pharmacia) equilibrated with 20 nM NaPO.sub.4, pH 6.0. The column was washed using an increasing NaCl gradient from 0 to 100 mM. The OspA-containing fractions were concentrated by ultrafiltration and stored at -70.degree. C.

As previously indicated, both lipidated and non-lipidated (truncated, i.e., without the first 17 amino acids) forms of OspC, OspA and OspC/OspA chimeric proteins were generated.

C. Immunization of Mice and Serologic Characterization Using ELISA (Enzyme-Linked Immunosorbent Assay)

Immunization of Mice

Mice, either C3H-J or ICR, were immunized with 3 ug of lipidated OspC/OspA chimeric protein or 6 ug of non-lipidated OspC/OspA chimers in 100 ul of aluminum hydroxide adjuvant (concentration of 1.8 mg/ml) by (SC) subcutaneous injection. As a negative control, mice were immunized with 100 ul of aluminum hydroxide adjuvant only. All mice received a total of three injections which were given at two week intervals. One week after the final immunization, blood was drawn from each mouse (including negative controls) and the serum was tested for IgG reactivity using the ELISA method described below, for the presence of anti-OspA antibodies to three different purified OspA proteins (Borrelia burgdorferi sensu stricto (B31), Borrelia garinii (K48) and Borrelia afzelli (PGau). The sera was tested at a dilution of 1:1000.

Mice were immunized with the chimeric proteins described in Table V.

TABLE-US-00016 TABLE V Chimeric Proteins Used to Immunize Mice SEQ ID SEQ ID NO.: NO.: Description (nucleic (polypep- FIG. Name (amino acid) acid) tide) No: OspA OspA-B31(18-273) 6 7 47, 48 OspC OspC-B31(19-211) 29 30 47, 48 OspC2- OspC-C2(19-204)/ 149 150 47, 48 OspA OspA-B31(18-273) .sup.1lipOspAP/Bo OspA-PGau(1-217)/ 139 140 49, 50 OspA-Bo(218-273) .sup.1lipOspAB/P OspA-B31(1-216)/ * * 49, 50 OspA-PKo(217-273) OspC- OspC-B31(19-211)/ 155 156 49, 50, OspAB/P OspA-B31(18-216)/ 53, 52, OspA-PKo(217-273) 54 OspCB31- OspC-B31(19-211)/ 145 146 51, 52, OspAB31 OspA-B31(18-273) 53, 54 OspC2- OspC-C2(19-204)/ 149 150 51, 52 OspAB31 OspA-B31(18-273) .sup.1lip OspA K/T OspA-K48(1-217)/ * * 52 OspA-Tro(218-273) .sup.1lip OspC-B31(1-211) 29 30 51 OspC-B31 OspCB31- OspC-B31(19-211)/ 177 178 53, 54 OspABPBP OspA-B31(30-150)/ OspA-PKo(151-179)/ OspA-B31(180-216) (190 N deletion)/ OspA-PKo(217-273) B31/B31/PKo .sup.1"lip" means the chimeric protein contains its native N-terminal lipidation signal

Serologic Characterization Using ELISA (Enzyme-Linked Immunosorbent Assay) Immobilization of Antigen onto ELISA Plates

A solution of purified recombinant OspC or OspA protein from each of the Borrelia burgdorferi strains B31 (Borrelia burgdorferi sensu stricto), K48 (Borrelia garinii) and PGau (Borrelia afzelli) was added to sodium phosphate buffer, pH 9.0, and was used to coat a commercial microwell plate (MaxiSorp.RTM., Nunc). The coating procedure was as follows: 100 .mu.l of a solution containing the appropriate OspA or OspC protein (made up at a concentration of 250 ng/ml in the following coating buffer: 100 mM Bis-Tris propane, pH 9.7) was added to each well of a microtiter plate which was incubated for one hour at 37.degree. C. The antigen solution was removed from the wells, the plate was washed three times with phosphate buffered saline (PBS) pH 9.0, and 300 .mu.l of blocking buffer solution was added (3% dry milk, 0.1% polyoxyethylenesorbitan (referred to herein as Tween 20.TM.), 0.02% NaN.sub.3 in 100 nM Bis-Tris propane, pH 9.7). Following a one hour incubation at 37.degree. C., the plates were washed four times with TBS-Tween 20.TM. wash buffer (20 mM Tris-Cl, pH 7.5, 136 mM NaCl, 0.1% Tween 20.TM. and 0.02% NaN.sub.3) and then were allowed to dry. The plates were then wrapped in plastic and stored at 4.degree. C. until they were used.

ELISA (Enzyme-Linked Immunosorbent Assay) Tests

The standard procedure for the ELISA tests was as follows: mouse serum was diluted 1:1000 in sample dilution buffer (1% dry milk, 136 mM NaCl, 0.1% Tween 20.TM., 0.02% NaN.sub.3 in 20 mM Tris-Cl, pH7.5) and 100 .mu.l of the diluted serum was added to the ELISA microtiter plate wells that had been coated with antigen as described above. Following incubation for 1 hour at 37.degree. C., the samples were removed and the plates were washed four times in TBS-Tween.TM. (20 mM Tris-Cl, pH 7.5; 136 mM NaCl; 0.1% Tween 20.TM. and 0.02% NaN.sub.3). For the secondary antibody, goat anti-mouse antisera conjugated to alkaline phosphatase-specific for either IgM (Fc) or IgG (Fab), (Jackson Immuno Research Laboratories) was diluted 1:750 in sample dilution buffer (1% dry milk, 136 mM NaCl, 0.1% Tween 20.TM., 0.02% NaN.sub.3 in 20 mM Tris-Cl, pH7.5) and 100 .mu.l of the diluted secondary antibody was added to each well. Following incubation for thirty minutes at 37.degree. C., the plates were washed three times with TBS-Tween.TM. (20 mM Tris-Cl, pH 7.5; 136 mM NaCl; 0.1% Tween 20.TM. and 0.02% NaN.sub.3) and 100 .mu.l of Phosphatase Substrate solution (5 mg of p-nitrophenylphosphate tablets dissolved in 1.times. diethanolamine substrate buffer to yield a 2 mg/ml solution--Kirkegaard Perry Laboratory) was added to each well. The plates were incubated for thirty minutes at 37.degree. C. and 100 .mu.l of stop solution (5% EDTA) was added to each well. The absorbance at 405 nm was read on a microplate reader (Dynatech). A sample was considered positive if it produced an average absorbance greater than the mean of the negative controls plus three standard deviations.

Previous work has demonstrated that it is the carboxy-terminal region of OspA that contains the antigenic sites that provide the immunoprotective response. Thus, in addition to the ELISA test described above, a modified ELISA was performed (herein referred to as the Protective ELISA Test), wherein the purified N-terminal region of B31 OspA (amino acids 18-139) was used to block any antibodies present in the mouse serum that had specificity to this N-terminal OspA region. These protective ELISA tests were performed as above, except that 80 .mu.g/ml of a purified B31 OspA fragment (amino acids 18-139) was added to the diluted mouse serum prior to adding the sera to the antigen-coated ELISA microtiter plate wells.

Results of ELISA Tests

Using the above-described ELISA tests, it was demonstrated that mice immunized with a non-lipidated OspC/OspA chimeric protein (OspC2-OspA-composed of OspC (a.a. 19-204 from strain C2)/OspA (a.a. 18-273 from strain B31) (SEQ ID NO.150) produced an immune response both to OspA and OspC that was comparable to the immune response generated to non-lipidated OspA (OspA--a.a. 18-273 from strain B31) and non-lipidated OspC (OspC--a.a. 19-211 from strain B31) control proteins (FIG. 47). As indicated in FIG. 47 and described above, mice were immunized with OspA, OspC or OspC2-OspA proteins and immune responses of the sera were measured against B31 OspA antigen (stippled bars) and B31 OspC antigen (solid bars).

Using the above-described Protective ELISA Test, it was also shown that mice immunized with the same non-lipidated OspC/OspA chimeric protein (OspC2-OspA-composed of OspC (a.a. 19-204 from strain C2)/OspA (a.a. 18-273 from strain B31) (SEQ ID NO.150) produced an immune response to the C-terminal portion of OspA that was comparable to the immune response generated to the C-terminal portion of a non-lipidated OspA (OspA--a.a. 18-273 from strain B31) control protein (FIG. 48). As indicated in FIG. 48, mice were immunized with OspA, OspC or OspC2-OspA proteins and immune responses of the sera were measured against B31 OspA antigen. The protective antibody response to B31 OspA antigen is indicated in the stippled bars.

Thus, these results clearly demonstrate that non-lipidated chimeric OspC/OspA proteins are able to induce immune responses in mice that are comparable to the immune response generated against non-lipidated OspC and OspA control proteins.

It had been previously thought that the lipidation signals that are present on Borrelia burgdorferi outer surface proteins were required for immunogenicity and that OspC and OspA proteins that lacked this lipidation signal would be less or non-immunogenic. To test this idea, mice were immunized with a non-lipidated OspC/OspA chimeric protein (OspC-OspAB/P--composed of OspC (a.a. 19-211 from strain B31)/OspA (a.a. 18-216 from strain B31)/OspA (a.a. 217-273 from strain PKo)(SEQ ID NO: 156) as well as two lipidated OspA proteins, lipOspAP/Bo (composed of OspA (a.a. 1-217 from strain PGau)/OspA (a.a. 218-273 from strain Bo)) and lipOspAB/P (composed of OspA (a.a. 1-216 from strain B31)/OspA (a.a. 217-273 from strain PKo)) and were subjected ELISA tests. Mice immunized with the non-lipidated OspC/OspA chimeric protein (OspC-OspAB/P) produced an immune response to OspA from each of the Borrelia burgdorferi strains B31 (Borrelia burgdorferi sensu stricto), K48 (Borrelia garinii) and PGau (Borrelia afzelli), that was equivalent or greater than the immune response generated to the two lipidated OspA control proteins (lipOspAP/Bo and lipOspAb/P) (FIG. 49).

Similar results to these were obtained using the Protective ELISA Test described above. Mice immunized with the non-lipidated OspC/OspA chimeric protein (OspC-OspAB/P) produced an immune response to the C-terminal region of OspA from each of the Borrelia burgdorferi strains B31 (Borrelia burgdorferi sensu stricto), K48 (Borrelia garinii) and PGau (Borrelia afzelli), that was equivalent or greater than the immune response generated to the C-terminal region of OspA from the two lipidated OspA control proteins (lipOspAP/Bo and lipOspAb/P) (FIG. 50).

In addition to the comparisons between non-lipidated OspC/OspA chimeric proteins and lipidated OspA control proteins, experiments were also performed to compare non-lipidated OspC/OspA chimeric proteins with a lipidated OspC control protein (FIG. 51). Mice that were immunized with either the non-lipidated OspC/OspA chimeric protein OspCB31-OspAB31 (composed of OspC (a.a. 19-211 from strain B31)/OspA (a.a. 18-273 from strain B31) (SEQ ID NO:146) or the non-lipidated OspC/OspA chimeric protein OspC2-OspAB31 (composed of OspC (a.a. 19-204 from strain C2)/OspA (a.a. 18-273 from strain B31) (SEQ ID NO:150) produced an immune response to OspC derived from the Borrelia burgdorferi strain B31 that was comparable to the immune response produced by a lipidated OspC control protein (lip OspC-B31--composed of OspC (a.a. 1-211 from strain B31)) (FIG. 51).

Thus, these results clearly demonstrate that non-lipidated chimeric OspC/OspA proteins are able to induce immune responses against OspA and OspC that are comparable to the immune response generated against OspA and OspC using lipidated OspA or OspC control proteins. The use of unlipidated forms of these proteins as vaccine immunogens or diagnostic antigens is highly desirable because the product yield is much greater and the proteins are much easier to purify. For these reasons, the production of these proteins less expensive.

The OspC/OspA chimeric proteins of the present invention are also able to generate immune responses against OspA proteins that are derived from strains that are not represented in the chimeric protein. Mice immunized with the OspC/OspA chimeric proteins, OspCB31-OspAB31 (SEQ ID NO: 146) and OspC2-OspAB31 (SEQ ID NO: 150), are not only able to generate immune responses that recognize OspA derived from strain B31 (Borrelia burgdorferi sensu stricto), but also recognize OspA derived from strain K48 (Borrelia garinii) and strain PGau (Borrelia afzelli) (FIG. 52). For comparison, mice were also immunized with the lipidated OspA chimeric protein, Lip OspA K/T (composed of OspA (a.a. 1-217 from strain K48)/OspA (a.a. 218-273 from strain Tro)) (FIG. 52).

Additional antibody responses to OspA derived from strain B31 (Borrelia burgdorferi sensu stricto), strain K48 (Borrelia garinii) and strain PGau (Borrelia afzelli) are also presented for sera from mice immunized with other OspC/OspA chimeric proteins. Thus, FIG. 53 presents the ELISA results from mice immunized with either OspCB31-OspAB/P (SEQ ID NO: 156), OspCB31-OspABPBP (SEQ ID NO: 178) or OspCB31-OspAB31 (SEQ ID NO: 146). In each case, sera from the immunized mice was tested against OspA derived from each of strain B31 (Borrelia burgdorferi sensu stricto), K48 (Borrelia garinii) and PGau (Borrelia afzelli). In all cases, a strong immune response was generated (FIG. 53). As with the previously described OspC/OspA chimeric proteins, the three OspC/OspA chimeric proteins used to immunize the mice in FIG. 52 also elicited a strong immune response to the C-terminal region of OspA when examined using the Protective ELISA Test described above (FIG. 54).

Tick Challenge of Immunized Mice

Mice, either C3H-J or JCR, that had been immunized as described above, were also challenged with either laboratory-infected nympha or field nympha. The immunized mice were placed in isolation cages and each mouse received 5-10 nymphs. All of the nymphs were collected and counted after 6 days. Four weeks after challenge, the mice were bled and sera was tested using commercially-available Western blot strips to Borrelia burgdorferi sensu stricto strain B331 (MarDx strips) and/or Borrelia garinii (MRL strips). Eight weeks after challenge, the mice were bled, sera was tested again by Western blot and ear punch and bladder samples were cultured. As a positive control, mice which had been immunized with only aluminum hydroxide adjuvant, as described above, were subjected to the same challenge.

The results of the tick challenge studies (Table VI) demonstrate that while immunization with lipidated OspC protein was unable to protect the mice, as evidenced by a positive Western blot signal (in 4 out of 5 mice), immunization with two different OspC/OspA chimeric proteins (SEQ ID NO.146 and SEQ ID NO.150) did provide protection, as indicated by the absence of Western blot signal (in 0 out of 8 mice and 0 out of 3 mice) (Table VI). The sham positive control showed that the challenge by the ticks was successful in all cases, as evidenced by 100% positive signal in Western blots (Table VI).

TABLE-US-00017 TABLE VI Effect of Vaccination on Transmission of Borrelia from Ticks Seroconversion Seroconversion Vaccine (Western Blots) (Western Blots) Candidate Mouse Tick-nymph Vaccinated Sham OspC1- C3H-J Long Island 0+/8 8+/8 OspAB31 OspC2- C3H-J Long Island 0+/3 4+/4 OspAB31 Lip OspC12 ICR Long Island 4+/5 5+/5

While this invention has been particularly shown and described with references to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims.

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2 DNA Artificial Sequence Oligonucleotide Primer tgact ctgacactag tgc 23 2 3rtificial Sequence Oligonucleotide Primer 2 gctactaaaa aaaccgggaa atggaattca 3DNA Artificial Sequence Oligonucleotide Primer 3 gcagcttggg attcaaaaac atccacttta aca 33 4 Artificial Sequence Oligonucleotide Primer 4 ggagaatata ttatgaaa DNA Artificial Sequence Oligonucleotide Primer 5 ctccttattt taaagcg 2 DNA Borrelia burgdorferi CDS (22) 6 atg aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gac gag aaa aac agc gtt tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 gat ttg cct ggt gaa atg aaa gtt ctt gta agc aaa gaa aaa aac aaa Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys 35 4c ggc aag tac gat cta att gca aca gta gac aag ctt gag ctt aaa Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys 5 gga act tct gat aaa aac aat gga tct gga gta ctt gaa ggc gta aaa 24hr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 gct gac aaa agt aaa gta aaa tta aca att tct gac gat cta ggt caa 288 Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln 85 9c aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca cta gta tca aaa 336 Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys gta act tcc aaa gac aag tca tca aca gaa gaa aaa ttc aat gaa 384 Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu ggt gaa gta tct gaa aaa ata ata aca aga gca gac gga acc aga 432 Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg gaa tac aca gga att aaa agc gat gga tct gga aaa gct aaa gag 48lu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu gtt tta aaa ggc tat gtt ctt gaa gga act cta act gct gaa aaa aca 528 Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr ttg gtg gtt aaa gaa gga act gtt act tta agc aaa aat att tca 576 Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser tct ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt gct 624 Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2act aaa aaa act gca gct tgg aat tca ggc act tca act tta aca 672 Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Gly Thr Ser Thr Leu Thr 222ct gta aac agt aaa aaa act aaa gac ctt gtg ttt aca aaa gaa 72hr Val Asn Ser Lys Lys Thr Lys Asp Leu Val Phe Thr Lys Glu 225 234ca att aca gta caa caa tac gac tca aat ggc acc aaa tta gag 768 Asn Thr Ile Thr Val Gln Gln Tyr Asp Ser Asn Gly Thr Lys Leu Glu 245 25gg tca gca gtt gaa att aca aaa ctt gat gaa att aaa aac gct tta 8Ser Ala Val Glu Ile Thr Lys Leu Asp Glu Ile Lys Asn Ala Leu 267aa 822 Lys * 7 273 PRT Borrelia burgdorferi 7 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys 35 4p Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys 5 Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln 85 9r Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2Thr Lys Lys Thr Ala Ala Trp Asn Ser Gly Thr Ser Thr Leu Thr 222hr Val Asn Ser Lys Lys Thr Lys Asp Leu Val Phe Thr Lys Glu 225 234hr Ile Thr Val Gln Gln Tyr Asp Ser Asn Gly Thr Lys Leu Glu 245 25ly Ser Ala Val Glu Ile Thr Lys Leu Asp Glu Ile Lys Asn Ala Leu 267 825 DNA Borrelia burgdorferi CDS (25) 8 atg aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gat gaa aaa aat agc gtt tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 gat tta cct ggt gga atg aca gtt ctt gta agt aaa gaa aaa gac aaa Leu Pro Gly Gly Met Thr Val Leu Val Ser Lys Glu Lys Asp Lys 35 4c ggt aaa tac agt cta gag gca aca gta gac aag ctt gag ctt aaa Gly Lys Tyr Ser Leu Glu Ala Thr Val Asp Lys Leu Glu Leu Lys 5 gga act tct gat aaa aac aac ggt tct gga aca ctt gaa ggt gaa aaa 24hr Ser Asp Lys Asn Asn Gly Ser Gly Thr Leu Glu Gly Glu Lys 65 7 act gac aaa agt aaa gta aaa tta aca att gct gat gac cta agt caa 288 Thr Asp Lys Ser Lys Val Lys Leu Thr Ile Ala Asp Asp Leu Ser Gln 85 9t aaa ttt gaa att ttc aaa gaa gat gcc aaa aca tta gta tca aaa 336 Thr Lys Phe Glu Ile Phe Lys Glu Asp Ala Lys Thr Leu Val Ser Lys gta acc ctt aaa gac aag tca tca aca gaa gaa aaa ttc aac gaa 384 Lys Val Thr Leu Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu ggt gaa aca tct gaa aaa aca ata gta aga gca aat gga acc aga 432 Lys Gly Glu Thr Ser Glu Lys Thr Ile Val Arg Ala Asn Gly Thr Arg gaa tac aca gac ata aaa agc gat gga tcc gga aaa gct aaa gaa 48lu Tyr Thr Asp Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu gtt tta aaa gac ttt act ctt gaa gga act cta gct gct gac ggc aaa 528 Val Leu Lys Asp Phe Thr Leu Glu Gly Thr Leu Ala Ala Asp Gly Lys aca ttg aaa gtt aca gaa ggc act gtt gtt tta agc aag aac att 576 Thr Thr Leu Lys Val Thr Glu Gly Thr Val Val Leu Ser Lys Asn Ile aaa tcc gga gaa ata aca gtt gca ctt gat gac tct gac act act 624 Leu Lys Ser Gly Glu Ile Thr Val Ala Leu Asp Asp Ser Asp Thr Thr 2gct act aaa aaa act gga aaa tgg gat tca aaa act tcc act tta 672 Gln Ala Thr Lys Lys Thr Gly Lys Trp Asp Ser Lys Thr Ser Thr Leu 222tt agt gtg aat agc caa aaa acc aaa aac ctt gta ttc aca aaa 72le Ser Val Asn Ser Gln Lys Thr Lys Asn Leu Val Phe Thr Lys 225 234ac aca ata aca gta caa aaa tac gac tca gca ggc acc aat cta 768 Glu Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu 245 25aa ggc aaa gca gtc gaa att aca aca ctt aaa gaa ctt aaa aac gct 8Gly Lys Ala Val Glu Ile Thr Thr Leu Lys Glu Leu Lys Asn Ala 267aa taa 825 Leu Lys * 9 274 PRT Borrelia burgdorferi 9 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 Asp Leu Pro Gly Gly Met Thr Val Leu Val Ser Lys Glu Lys Asp Lys 35 4p Gly Lys Tyr Ser Leu Glu Ala Thr Val Asp Lys Leu Glu Leu Lys 5 Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Thr Leu Glu Gly Glu Lys 65 7 Thr Asp Lys Ser Lys Val Lys Leu Thr Ile Ala Asp Asp Leu Ser Gln 85 9r Lys Phe Glu Ile Phe Lys Glu Asp Ala Lys Thr Leu Val Ser Lys Val Thr Leu Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Gly Glu Thr Ser Glu Lys Thr Ile Val Arg Ala Asn Gly Thr Arg Glu Tyr Thr Asp Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys Asp Phe Thr Leu Glu Gly Thr Leu Ala Ala Asp Gly Lys Thr Leu Lys Val Thr Glu Gly Thr Val Val Leu Ser Lys Asn Ile Lys Ser Gly Glu Ile Thr Val Ala Leu Asp Asp Ser Asp Thr Thr 2Ala Thr Lys Lys Thr Gly Lys Trp Asp Ser Lys Thr Ser Thr Leu 222le Ser Val Asn Ser Gln Lys Thr Lys Asn Leu Val Phe Thr Lys 225 234sp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu 245 25lu Gly Lys Ala Val Glu Ile Thr Thr Leu Lys Glu Leu Lys Asn Ala 267ys DNA Borrelia burgdorferi CDS (22) aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gat gaa aaa aac agc gct tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Ala Ser Val 2 gat ttg cct ggt gag atg aaa gtt ctt gta agt aaa gaa aaa gac aaa Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asp Lys 35 4c ggt aag tac agt cta aag gca aca gta gac aag att gag cta aaa Gly Lys Tyr Ser Leu Lys Ala Thr Val Asp Lys Ile Glu Leu Lys 5 gga act tct gat aaa gac aat ggt tct gga gtg ctt gaa ggt aca aaa 24hr Ser Asp Lys Asp Asn Gly Ser Gly Val Leu Glu Gly Thr Lys 65 7 gat gac aaa agt aaa gca aaa tta aca att gct gac gat cta agt aaa 288 Asp Asp Lys Ser Lys Ala Lys Leu Thr Ile Ala Asp Asp Leu Ser Lys 85 9c aca ttc gaa ctt tta aaa gaa gat ggc aaa aca tta gtg tca aga 336 Thr Thr Phe Glu Leu Leu Lys Glu Asp Gly Lys Thr Leu Val Ser Arg gta agt tct aga gac aaa aca tca aca gat gaa atg ttc aat gaa 384 Lys Val Ser Ser Arg Asp Lys Thr Ser Thr Asp Glu Met Phe Asn Glu ggt gaa ttg tct gca aaa acc atg aca aga gaa aat gga acc aaa 432 Lys Gly Glu Leu Ser Ala Lys Thr Met Thr Arg Glu Asn Gly Thr Lys gaa tat aca gaa atg aaa agc gat gga acc gga aaa gct aaa gaa 48lu Tyr Thr Glu Met Lys Ser Asp Gly Thr Gly Lys Ala Lys Glu gtt tta aaa aag ttt act ctt gaa gga aaa gta gct aat gat aaa gta 528 Val Leu Lys Lys Phe Thr Leu Glu Gly Lys Val Ala Asn Asp Lys Val ttg gaa gta aaa gaa gga acc gtt act tta agt aag gaa att gca 576 Thr Leu Glu Val Lys Glu Gly Thr Val Thr Leu Ser Lys Glu Ile Ala tct gga gaa gta aca gtt gct ctt aat gac act aac act act cag 624 Lys Ser Gly Glu Val Thr Val Ala Leu Asn Asp Thr Asn Thr Thr Gln 2act aaa aaa act ggc gca tgg gat tca aaa act tct act tta aca 672 Ala Thr Lys Lys Thr Gly Ala Trp Asp Ser Lys Thr Ser Thr Leu Thr 222gt gtt aac agc aaa aaa act aca caa ctt gtg ttt act aaa caa 72er Val Asn Ser Lys Lys Thr Thr Gln Leu Val Phe Thr Lys Gln 225 234ca ata act gta aaa caa tac gac tcc gca ggt acc aat tta gaa 768 Tyr Thr Ile Thr Val Lys Gln Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25gc aca gca gtc gaa att aaa aca ctt gat gaa ctt aaa aac gct tta 8Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu 267aa 822 Lys * PRT Borrelia burgdorferi Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Ala Ser Val 2 Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asp Lys 35 4p Gly Lys Tyr Ser Leu Lys Ala Thr Val Asp Lys Ile Glu Leu Lys 5 Gly Thr Ser Asp Lys Asp Asn Gly Ser Gly Val Leu Glu Gly Thr Lys 65 7 Asp Asp Lys Ser Lys Ala Lys Leu Thr Ile Ala Asp Asp Leu Ser Lys 85 9r Thr Phe Glu Leu Leu Lys Glu Asp Gly Lys Thr Leu Val Ser Arg Val Ser Ser Arg Asp Lys Thr Ser Thr Asp Glu Met Phe Asn Glu Gly Glu Leu Ser Ala Lys Thr Met Thr Arg Glu Asn Gly Thr Lys Glu Tyr Thr Glu Met Lys Ser Asp Gly Thr Gly Lys Ala Lys Glu Val Leu Lys Lys Phe Thr Leu Glu Gly Lys Val Ala Asn Asp Lys Val Leu Glu Val Lys Glu Gly Thr Val Thr Leu Ser Lys Glu Ile Ala Ser Gly Glu Val Thr Val Ala Leu Asn Asp Thr Asn Thr Thr Gln 2Thr Lys Lys Thr Gly Ala Trp Asp Ser Lys Thr Ser Thr Leu Thr 222er Val Asn Ser Lys Lys Thr Thr Gln Leu Val Phe Thr Lys Gln 225 234hr Ile Thr Val Lys Gln Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25ly Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu 2672 8Borrelia burgdorferi CDS (atg aaa aaa tat tta ttg gga ata ggt cta ata tta gct tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gac gag aaa aac agc gtt tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 gat ttg cct ggt gaa atg aaa gtt ctt gta agc aaa gaa aaa gac aaa Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asp Lys 35 4c ggc aag tac agt cta atg gca aca gta gac aag ctt gag ctt aaa Gly Lys Tyr Ser Leu Met Ala Thr Val Asp Lys Leu Glu Leu Lys 5 gga aca tct gat aaa aac aat gga tct ggg gtg ctt gaa ggc gta aaa 24hr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 gct gac aaa agc aaa gta aaa tta aca gtt tct gac gat cta agc aca 288 Ala Asp Lys Ser Lys Val Lys Leu Thr Val Ser Asp Asp Leu Ser Thr 85

9c aca ctt gaa gtt tta aaa gaa gat ggc aaa aca tta gtg tca aaa 336 Thr Thr Leu Glu Val Leu Lys Glu Asp Gly Lys Thr Leu Val Ser Lys aga act tct aaa gat aag tca tca aca gaa gaa aag ttc aat gaa 384 Lys Arg Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu ggc gaa tta gtt gaa aaa ata atg gca aga gca aac gga acc ata 432 Lys Gly Glu Leu Val Glu Lys Ile Met Ala Arg Ala Asn Gly Thr Ile gaa tac aca gga att aaa agc gat gga tcc gga aaa gct aaa gaa 48lu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu act tta aaa gaa tat gtt ctt gaa gga act cta act gct gaa aaa gca 528 Thr Leu Lys Glu Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Ala ttg gtg gtt aaa gaa gga act gtt act tta agt aag cac att tca 576 Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys His Ile Ser tct gga gaa gta aca gct gaa ctt aat gac act gac agt act caa 624 Lys Ser Gly Glu Val Thr Ala Glu Leu Asn Asp Thr Asp Ser Thr Gln 2act aaa aaa act ggg aaa tgg gat gca ggc act tca act tta aca 672 Ala Thr Lys Lys Thr Gly Lys Trp Asp Ala Gly Thr Ser Thr Leu Thr 222ct gta aac aac aaa aaa act aaa gcc ctt gta ttt aca aaa caa 72hr Val Asn Asn Lys Lys Thr Lys Ala Leu Val Phe Thr Lys Gln 225 234ca att aca tca caa aaa tac gac tca gca gga acc aac ttg gaa 768 Asp Thr Ile Thr Ser Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25gc aca gca gtc gaa att aaa aca ctt gat gaa ctt aaa aac gct tta 8Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu 267PRT Borrelia burgdorferi Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asp Lys 35 4p Gly Lys Tyr Ser Leu Met Ala Thr Val Asp Lys Leu Glu Leu Lys 5 Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 Ala Asp Lys Ser Lys Val Lys Leu Thr Val Ser Asp Asp Leu Ser Thr 85 9r Thr Leu Glu Val Leu Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Arg Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Gly Glu Leu Val Glu Lys Ile Met Ala Arg Ala Asn Gly Thr Ile Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Thr Leu Lys Glu Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Ala Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys His Ile Ser Ser Gly Glu Val Thr Ala Glu Leu Asn Asp Thr Asp Ser Thr Gln 2Thr Lys Lys Thr Gly Lys Trp Asp Ala Gly Thr Ser Thr Leu Thr 222hr Val Asn Asn Lys Lys Thr Lys Ala Leu Val Phe Thr Lys Gln 225 234hr Ile Thr Ser Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25ly Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu 2674 2rtificial Sequence Oligonucleotide Primer gcaaaa accatgacaa g 2 DNA Artificial Sequence Oligonucleotide Primer tcaaca gaagaaaaat tc 22 NA Artificial Sequence Oligonucleotide Primer atccat atgaaaaaat atttattggg 3 DNA Artificial Sequence Oligonucleotide Primer gatcca tatggctaag caaaatgtta gc 32 NA Artificial Sequence Oligonucleotide Primer tcaagt actccaga 7 DNA Artificial Sequence Oligonucleotide Primer tctaga tcttatttta aagcgtt 27 2A Artificial Sequence Oligonucleotide Primer 2cggtg accttttaaa gcgtttttaa t 3orrelia burgdorferi CDS (9tg aga tta tta ata gga ttt gct tta gcg tta gct tta ata gga tgt 48 Met Arg Leu Leu Ile Gly Phe Ala Leu Ala Leu Ala Leu Ile Gly Cys caa aaa ggt gct gag tca att ggt tct caa aaa gaa aat gat cta 96 Ala Gln Lys Gly Ala Glu Ser Ile Gly Ser Gln Lys Glu Asn Asp Leu 2 aac ctt gaa gac tct agt aaa aaa tca cat caa aac gct aaa caa gac Leu Glu Asp Ser Ser Lys Lys Ser His Gln Asn Ala Lys Gln Asp 35 4t cct gcg gtg aca gaa gac tca gtg tct ttg ttt aat ggt aat aaa Pro Ala Val Thr Glu Asp Ser Val Ser Leu Phe Asn Gly Asn Lys 5 att ttt gta agc aaa gaa aaa aat agc tcc ggc aaa tat gat tta aga 24he Val Ser Lys Glu Lys Asn Ser Ser Gly Lys Tyr Asp Leu Arg 65 7 gca aca att gat cag gtt gaa ctt aaa gga act tcc gat aaa aac aat 288 Ala Thr Ile Asp Gln Val Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn 85 9t tct gga acc ctt gaa ggt tca aag cct gac aag agt aaa gta aaa 336 Gly Ser Gly Thr Leu Glu Gly Ser Lys Pro Asp Lys Ser Lys Val Lys aca gtt tct gct gat tta aac aca gta acc tta gaa gca ttt gat 384 Leu Thr Val Ser Ala Asp Leu Asn Thr Val Thr Leu Glu Ala Phe Asp agc aac caa aaa att tca agt aaa gtt act aaa aaa cag ggg tca 432 Ala Ser Asn Gln Lys Ile Ser Ser Lys Val Thr Lys Lys Gln Gly Ser aca gag gaa act ctc aaa gct aat aaa tta gac tca aag aaa tta 48hr Glu Glu Thr Leu Lys Ala Asn Lys Leu Asp Ser Lys Lys Leu aca aga tca aac gga act aca ctt gaa tac tca caa ata aca gat gct 528 Thr Arg Ser Asn Gly Thr Thr Leu Glu Tyr Ser Gln Ile Thr Asp Ala aat gct aca aaa gca gta gaa act cta aaa aat agc att aag ctt 576 Asp Asn Ala Thr Lys Ala Val Glu Thr Leu Lys Asn Ser Ile Lys Leu gga agt ctt gta gtc gga aaa aca aca gtg gaa att aaa gaa ggt 624 Glu Gly Ser Leu Val Val Gly Lys Thr Thr Val Glu Ile Lys Glu Gly 2gtt act cta aaa aga gaa att gaa aaa gat gga aaa gta aaa gtc 672 Thr Val Thr Leu Lys Arg Glu Ile Glu Lys Asp Gly Lys Val Lys Val 222tg aat gac act gca ggt tct aac aaa aaa aca ggt aaa tgg gaa 72eu Asn Asp Thr Ala Gly Ser Asn Lys Lys Thr Gly Lys Trp Glu 225 234gt act agc act tta aca att agt gct gac agc aaa aaa act aaa 768 Asp Ser Thr Ser Thr Leu Thr Ile Ser Ala Asp Ser Lys Lys Thr Lys 245 25at ttg gtg ttc tta aca gat ggt aca att aca gta caa caa tac aac 8Leu Val Phe Leu Thr Asp Gly Thr Ile Thr Val Gln Gln Tyr Asn 267ct gga acc agc cta gaa gga tca gca agt gaa att aaa aat ctt 864 Thr Ala Gly Thr Ser Leu Glu Gly Ser Ala Ser Glu Ile Lys Asn Leu 275 28ca gag ctt aaa aac gct tta aaa taa 89lu Leu Lys Asn Ala Leu Lys * 292 296 PRT Borrelia burgdorferi 22 Met Arg Leu Leu Ile Gly Phe Ala Leu Ala Leu Ala Leu Ile Gly Cys Gln Lys Gly Ala Glu Ser Ile Gly Ser Gln Lys Glu Asn Asp Leu 2 Asn Leu Glu Asp Ser Ser Lys Lys Ser His Gln Asn Ala Lys Gln Asp 35 4u Pro Ala Val Thr Glu Asp Ser Val Ser Leu Phe Asn Gly Asn Lys 5 Ile Phe Val Ser Lys Glu Lys Asn Ser Ser Gly Lys Tyr Asp Leu Arg 65 7 Ala Thr Ile Asp Gln Val Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn 85 9y Ser Gly Thr Leu Glu Gly Ser Lys Pro Asp Lys Ser Lys Val Lys Thr Val Ser Ala Asp Leu Asn Thr Val Thr Leu Glu Ala Phe Asp Ser Asn Gln Lys Ile Ser Ser Lys Val Thr Lys Lys Gln Gly Ser Thr Glu Glu Thr Leu Lys Ala Asn Lys Leu Asp Ser Lys Lys Leu Thr Arg Ser Asn Gly Thr Thr Leu Glu Tyr Ser Gln Ile Thr Asp Ala Asn Ala Thr Lys Ala Val Glu Thr Leu Lys Asn Ser Ile Lys Leu Gly Ser Leu Val Val Gly Lys Thr Thr Val Glu Ile Lys Glu Gly 2Val Thr Leu Lys Arg Glu Ile Glu Lys Asp Gly Lys Val Lys Val 222eu Asn Asp Thr Ala Gly Ser Asn Lys Lys Thr Gly Lys Trp Glu 225 234er Thr Ser Thr Leu Thr Ile Ser Ala Asp Ser Lys Lys Thr Lys 245 25sp Leu Val Phe Leu Thr Asp Gly Thr Ile Thr Val Gln Gln Tyr Asn 267la Gly Thr Ser Leu Glu Gly Ser Ala Ser Glu Ile Lys Asn Leu 275 28er Glu Leu Lys Asn Ala Leu Lys 293 Artificial Sequence Oligonucleotide Primer 23 ggtacaatta cagtacaa 2 DNA Artificial Sequence Oligonucleotide Primer 24 ccgagaatct catatggcac aaaaaggtgc tgagtcaatt gg 42 25 35 DNA Artificial Sequence Oligonucleotide Primer 25 ccgatatcgg atcctatttt aaagcgtttt taagc 35 26 3rtificial Sequence Oligonucleotide Primer 26 ggatccggtg accttttaaa gcgtttttaa g 3 DNA Artificial Sequence Oligonucleotide Primer 27 gtgcgcgacc atatgaaaaa gaatacatta agtgcg 36 28 36 DNA Artificial Sequence Oligonucleotide Primer 28 gtcggcggat ccttaaggtt tttttggact ttctgc 36 29 633 DNA Borrelia burgdorferi CDS (33) 29 atg aaa aag aat aca tta agt gcg ata tta atg act tta ttt tta ttt 48 Met Lys Lys Asn Thr Leu Ser Ala Ile Leu Met Thr Leu Phe Leu Phe tct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 96 Ile Ser Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser 2 gct gat gag tct gtt aaa ggg cct aat ctt aca gaa ata agt aaa aaa Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 35 4t acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 5 ttg ctg tca tct ata gat gaa att gct gct aaa gct att ggt aaa aaa 24eu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 65 7 ata cac caa aat aat ggt ttg gat acc gaa tat aat cac aat gga tca 288 Ile His Gln Asn Asn Gly Leu Asp Thr Glu Tyr Asn His Asn Gly Ser 85 9g tta gcg gga cgt tat gca ata tca acc cta ata aaa caa aaa tta 336 Leu Leu Ala Gly Arg Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu gga ttg aaa aat gaa gga tta aag gaa aaa att gat gcg gct aag 384 Asp Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys tgt tct gaa aca ttt act aat aaa tta aaa gaa aaa cac aca gat 432 Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp ggt aaa gaa ggt gtt act gat gct gat gca aaa gaa gcc att tta 48ly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu aaa aca aat ggt act aaa act aaa ggt gct gaa gaa ctt gga aaa tta 528 Lys Thr Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu gaa tca gta gag gtc ttg tca aaa gca gct aaa gag atg ctt gct 576 Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala tca gtt aaa gag ctt aca agc cct gtt gtg gca gaa agt cca aaa 624 Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys 2cct taa 633 Lys Pro * 2Borrelia burgdorferi 3ys Lys Asn Thr Leu Ser Ala Ile Leu Met Thr Leu Phe Leu Phe Ser Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser 2 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 35 4e Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 5 Leu Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 65 7 Ile His Gln Asn Asn Gly Leu Asp Thr Glu Tyr Asn His Asn Gly Ser 85 9u Leu Ala Gly Arg Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu Lys Thr Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys 2Pro 23orrelia burgdorferi CDS (3tg aaa aag aat aca tta agt gcg ata tta atg act tta ttt tta ttt 48 Met Lys Lys Asn Thr Leu Ser Ala Ile Leu Met Thr Leu Phe Leu Phe tct tgt aat aat tca ggt ggg gat acc gca tct act aat cct gat 96 Ile Ser Cys Asn Asn Ser Gly Gly Asp Thr Ala Ser Thr Asn Pro Asp 2 gag tct gca aaa gga cct aat ctt aca gta ata agc aaa aaa att aca Ser Ala Lys Gly Pro Asn Leu Thr Val Ile Ser Lys Lys Ile Thr 35 4t tct aat gca ttt gta ctg gct gtg aaa gaa gtt gag gct ttg atc Ser Asn Ala Phe Val Leu Ala Val Lys Glu Val Glu Ala Leu Ile 5 tca tct ata gat gaa ctt gct aat aaa gct att ggt aaa gta ata cat 24er Ile Asp Glu Leu Ala Asn Lys Ala Ile Gly Lys Val Ile His 65 7 caa aat aat ggt tta aat gct aat gcg ggt caa aac gga tca ttg tta 288 Gln Asn Asn Gly Leu Asn Ala Asn Ala Gly Gln Asn Gly Ser Leu Leu 85 9a gga gcc tat gca ata tca acc cta ata aca gaa aaa tta agt aaa 336 Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Thr Glu Lys Leu Ser Lys aaa aat tca gaa gag tta aat aaa aaa att gaa gag gct aag aac 384 Leu Lys Asn Ser Glu Glu Leu Asn Lys Lys Ile Glu Glu Ala Lys Asn tct gaa gca ttt act aat aga cta aaa ggt tct cat gca caa ctt 432 His Ser Glu Ala Phe Thr Asn Arg Leu Lys Gly Ser His Ala Gln Leu gtt gct gct gct act gat gat cat gca aaa gaa gct att tta aag 48al Ala Ala Ala Thr Asp Asp His Ala Lys Glu Ala Ile Leu Lys tca aat cct act aaa gat aag ggt gct aaa gca ctt aaa gac tta tct 528 Ser Asn Pro Thr Lys Asp Lys Gly Ala Lys Ala Leu Lys Asp Leu Ser

tca gta gaa agc ttg gca aaa gca gcg caa gaa gca tta gct aat 576 Glu Ser Val Glu Ser Leu Ala Lys Ala Ala Gln Glu Ala Leu Ala Asn gtt aaa gaa ctt aca aat cct gtt gtg gca gaa agt cca aaa aaa 624 Ser Val Lys Glu Leu Thr Asn Pro Val Val Ala Glu Ser Pro Lys Lys 2taa 63 32 2Borrelia burgdorferi 32 Met Lys Lys Asn Thr Leu Ser Ala Ile Leu Met Thr Leu Phe Leu Phe Ser Cys Asn Asn Ser Gly Gly Asp Thr Ala Ser Thr Asn Pro Asp 2 Glu Ser Ala Lys Gly Pro Asn Leu Thr Val Ile Ser Lys Lys Ile Thr 35 4p Ser Asn Ala Phe Val Leu Ala Val Lys Glu Val Glu Ala Leu Ile 5 Ser Ser Ile Asp Glu Leu Ala Asn Lys Ala Ile Gly Lys Val Ile His 65 7 Gln Asn Asn Gly Leu Asn Ala Asn Ala Gly Gln Asn Gly Ser Leu Leu 85 9a Gly Ala Tyr Ala Ile Ser Thr Leu Ile Thr Glu Lys Leu Ser Lys Lys Asn Ser Glu Glu Leu Asn Lys Lys Ile Glu Glu Ala Lys Asn Ser Glu Ala Phe Thr Asn Arg Leu Lys Gly Ser His Ala Gln Leu Val Ala Ala Ala Thr Asp Asp His Ala Lys Glu Ala Ile Leu Lys Ser Asn Pro Thr Lys Asp Lys Gly Ala Lys Ala Leu Lys Asp Leu Ser Ser Val Glu Ser Leu Ala Lys Ala Ala Gln Glu Ala Leu Ala Asn Val Lys Glu Leu Thr Asn Pro Val Val Ala Glu Ser Pro Lys Lys 233 639 DNA Borrelia burgdorferi CDS (39) 33 atg aaa aag aat aca tta agt gcg ata tta atg act tta ttt tta ttt 48 Met Lys Lys Asn Thr Leu Ser Ala Ile Leu Met Thr Leu Phe Leu Phe tct tgt agt aat tca ggg aaa ggt ggg gat tct gca tct act aat 96 Ile Ser Cys Ser Asn Ser Gly Lys Gly Gly Asp Ser Ala Ser Thr Asn 2 cct gct gac gag tct gcg aaa ggg cct aat ctt aca gaa ata agc aaa Ala Asp Glu Ser Ala Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys 35 4a att aca gat tct aat gca ttt gta ctt gct gtt aaa gaa gtt gag Ile Thr Asp Ser Asn Ala Phe Val Leu Ala Val Lys Glu Val Glu 5 act ttg gtt tta tct ata gat gaa ctt gct aag aaa gct att ggt caa 24eu Val Leu Ser Ile Asp Glu Leu Ala Lys Lys Ala Ile Gly Gln 65 7 aaa ata gac aat aat aat ggt tta gct gct tta aat aat cag aat gga 288 Lys Ile Asp Asn Asn Asn Gly Leu Ala Ala Leu Asn Asn Gln Asn Gly 85 9g ttg tta gca gga gcc tat gca ata tca acc cta ata aca gaa aaa 336 Ser Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Thr Glu Lys agt aaa ttg aaa aat tta gaa gaa tta aag aca gaa att gca aag 384 Leu Ser Lys Leu Lys Asn Leu Glu Glu Leu Lys Thr Glu Ile Ala Lys aag aaa tgt tcc gaa gaa ttt act aat aaa cta aaa agt ggt cat 432 Ala Lys Lys Cys Ser Glu Glu Phe Thr Asn Lys Leu Lys Ser Gly His gat ctt ggc aaa cag gat gct acc gat gat cat gca aaa gca gct 48sp Leu Gly Lys Gln Asp Ala Thr Asp Asp His Ala Lys Ala Ala att tta aaa aca cat gca act acc gat aaa ggt gct aaa gaa ttt aaa 528 Ile Leu Lys Thr His Ala Thr Thr Asp Lys Gly Ala Lys Glu Phe Lys tta ttt gaa tca gta gaa ggt ttg tta aaa gca gct caa gta gca 576 Asp Leu Phe Glu Ser Val Glu Gly Leu Leu Lys Ala Ala Gln Val Ala act aat tca gtt aaa gaa ctt aca agt cct gtt gta gca gaa agt 624 Leu Thr Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser 2aaa aaa cct taa 639 Pro Lys Lys Pro * 2Borrelia burgdorferi 34 Met Lys Lys Asn Thr Leu Ser Ala Ile Leu Met Thr Leu Phe Leu Phe Ser Cys Ser Asn Ser Gly Lys Gly Gly Asp Ser Ala Ser Thr Asn 2 Pro Ala Asp Glu Ser Ala Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys 35 4s Ile Thr Asp Ser Asn Ala Phe Val Leu Ala Val Lys Glu Val Glu 5 Thr Leu Val Leu Ser Ile Asp Glu Leu Ala Lys Lys Ala Ile Gly Gln 65 7 Lys Ile Asp Asn Asn Asn Gly Leu Ala Ala Leu Asn Asn Gln Asn Gly 85 9r Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Thr Glu Lys Ser Lys Leu Lys Asn Leu Glu Glu Leu Lys Thr Glu Ile Ala Lys Lys Lys Cys Ser Glu Glu Phe Thr Asn Lys Leu Lys Ser Gly His Asp Leu Gly Lys Gln Asp Ala Thr Asp Asp His Ala Lys Ala Ala Ile Leu Lys Thr His Ala Thr Thr Asp Lys Gly Ala Lys Glu Phe Lys Leu Phe Glu Ser Val Glu Gly Leu Leu Lys Ala Ala Gln Val Ala Thr Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser 2Lys Lys Pro 224 DNA Borrelia burgdorferi CDS (24) 35 atg aaa aag aat aca tta agt gcg ata tta atg act tta ttt tta ttt 48 Met Lys Lys Asn Thr Leu Ser Ala Ile Leu Met Thr Leu Phe Leu Phe tct tgt aat aat tca ggt ggg gat tct gca tct act aat cct gat 96 Ile Ser Cys Asn Asn Ser Gly Gly Asp Ser Ala Ser Thr Asn Pro Asp 2 gag tct gca aaa gga cct aat ctt acc gta ata agc aaa aaa att aca Ser Ala Lys Gly Pro Asn Leu Thr Val Ile Ser Lys Lys Ile Thr 35 4t tct aat gca ttt tta ctg gct gtg aaa gaa gtt gag gct ttg ctt Ser Asn Ala Phe Leu Leu Ala Val Lys Glu Val Glu Ala Leu Leu 5 tca tct ata gat gaa ctt tct aaa gct att ggt aaa aaa ata aaa aat 24er Ile Asp Glu Leu Ser Lys Ala Ile Gly Lys Lys Ile Lys Asn 65 7 gat ggt act tta gat aac gaa gca aat cga aac gaa tca ttg ata gca 288 Asp Gly Thr Leu Asp Asn Glu Ala Asn Arg Asn Glu Ser Leu Ile Ala 85 9a gct tat gaa ata tca aaa cta ata aca caa aaa tta agt gta ttg 336 Gly Ala Tyr Glu Ile Ser Lys Leu Ile Thr Gln Lys Leu Ser Val Leu tca gaa gaa tta aag aaa aaa att aaa gag gct aag gat tgt tcc 384 Asn Ser Glu Glu Leu Lys Lys Lys Ile Lys Glu Ala Lys Asp Cys Ser aaa ttt act act aag cta aaa gat agt cat gca gag ctt ggt ata 432 Glu Lys Phe Thr Thr Lys Leu Lys Asp Ser His Ala Glu Leu Gly Ile agc gtt cag gat gat aat gca aaa aaa gct att tta aaa aca cat 48er Val Gln Asp Asp Asn Ala Lys Lys Ala Ile Leu Lys Thr His gga act aaa gac aag ggt gct aaa gaa ctt gaa gag tta ttt aaa tca 528 Gly Thr Lys Asp Lys Gly Ala Lys Glu Leu Glu Glu Leu Phe Lys Ser gaa agc ttg tca aaa gca gcg caa gca gca tta act aat tca gtt 576 Leu Glu Ser Leu Ser Lys Ala Ala Gln Ala Ala Leu Thr Asn Ser Val gag ctt aca aat cct gtt gtg gca gaa agt cca aaa aaa cct taa 624 Lys Glu Leu Thr Asn Pro Val Val Ala Glu Ser Pro Lys Lys Pro * 2Borrelia burgdorferi 36 Met Lys Lys Asn Thr Leu Ser Ala Ile Leu Met Thr Leu Phe Leu Phe Ser Cys Asn Asn Ser Gly Gly Asp Ser Ala Ser Thr Asn Pro Asp 2 Glu Ser Ala Lys Gly Pro Asn Leu Thr Val Ile Ser Lys Lys Ile Thr 35 4p Ser Asn Ala Phe Leu Leu Ala Val Lys Glu Val Glu Ala Leu Leu 5 Ser Ser Ile Asp Glu Leu Ser Lys Ala Ile Gly Lys Lys Ile Lys Asn 65 7 Asp Gly Thr Leu Asp Asn Glu Ala Asn Arg Asn Glu Ser Leu Ile Ala 85 9y Ala Tyr Glu Ile Ser Lys Leu Ile Thr Gln Lys Leu Ser Val Leu Ser Glu Glu Leu Lys Lys Lys Ile Lys Glu Ala Lys Asp Cys Ser Lys Phe Thr Thr Lys Leu Lys Asp Ser His Ala Glu Leu Gly Ile Ser Val Gln Asp Asp Asn Ala Lys Lys Ala Ile Leu Lys Thr His Gly Thr Lys Asp Lys Gly Ala Lys Glu Leu Glu Glu Leu Phe Lys Ser Glu Ser Leu Ser Lys Ala Ala Gln Ala Ala Leu Thr Asn Ser Val Glu Leu Thr Asn Pro Val Val Ala Glu Ser Pro Lys Lys Pro 26 DNA Artificial Sequence Oligonucleotide Primer 37 gtgcgcgacc atatggctaa taattcaggg aaagat 36 38 36 DNA Artificial Sequence Oligonucleotide Primer 38 gtgcgcgacc atatggctag taattcaggg aaaggt 36 39 36 DNA Artificial Sequence Oligonucleotide Primer 39 gtgcgcgacc atatggctaa taattcaggt ggggat 36 4A Artificial Sequence Oligonucleotide Primer 4aaaat tatttgaa 5 DNA Artificial Sequence Oligonucleotide Primer 4tcacc ccatgggaaa taattcaggg aaagg 35 42 Artificial Sequence Oligonucleotide Primer 42 tatagatgac agcaacgc rtificial Sequence Oligonucleotide Primer 43 ccggtgaccc catggtacca ggtttttttg gactttctgc 4 DNA Artificial Sequence Oligonucleotide Primer 44 ccggatccat atggttaaaa aaataatatt tatttc 36 45 37 DNA Artificial Sequence Oligonucleotide Primer 45 gatatctaga tctttaattg ctctgctcac tctcttc 37 46 34 DNA Artificial Sequence Oligonucleotide Primer 46 ccgggatcca tatggctagt gcaattggtc gtgg 34 47 Artificial Sequence Oligonucleotide Primer 47 atgattatca atcataat 8 DNA Artificial Sequence Oligonucleotide Primer 48 tctgaacaat gacaaaac 25 DNA Borrelia burgdorferi 49 atgaaaaaat atttattggg aataggtcta atattagcct taatagcatg taagcaaaat 6cagcc ttgatgaaaa aaatagcgtt tcagtagatt tacctggtgg aatgacagtt gtaagta aagaaaaaga caaagacggt aaatacagtc tagaggcaac agtagacaag gagctta aaggaacttc tgataaaaac aacggttctg gaacacttga aggtgaaaaa 24caaaa gtaaagtaaa atcaacaatt gctgatgacc taagtcaaac taaatttgaa 3tcaaag aagatggcaa aacattagta tcaaaaaaag taacccttaa agacaagtca 36agaag aaaaattcaa cggaaagggt gaaacatctg aaaaaacaat agtaagagca 42aacca gacttgaata cacagacata aaaagcgatg gatccggaaa agctaaagaa 48aaaag actttactct tgaaggaact ctagctgctg acggcaaaac aacattgaaa 54agaag gcactgttgt tttaagcaag aacattttaa aatccggaga aataacagct 6ttgatg actctgacac tactcgggct actaaaaaaa ctggaaaatg ggattcaaag 66cactt taacaattag tgtgaatagc caaaaaacca aaaaccttgt attcacaaaa 72cacaa taacagtaca aagatacgac tcagcaggca ccaatctaga aggcaaagca 78aatta caacacttaa agaacttaaa aacgctttaa aataa 825 5NA Borrelia burgdorferi 5aaaat atttattggg aataggtcta atattagcat taatagcatg taagcaaaat 6cagcc ttgatgaaaa aaatagcgtt tcagtagatt tacctggtgg aatgcaagtt gtaagta aagaaaaaga caaagatggt aaatacagtc taatggcaac agtagacaag gagctta aaggaacttc tgataaaaac aacggttctg gaacacttga aggtgaaaaa 24caaaa gtaaagcaaa attaacaatt gctgaggatc taagtaaaac cacatttgaa 3tcaaag aagatggcaa aacattagta tcaaaaaaag taacccttaa agacaagtca 36agaag aaaaattcaa cgcaaagggt gaagcatctg aaaaaacaat agtaagagca 42aacca gacttgaata cacagacata aaaagcgata aaaccggaaa agctaaagaa 48aaaag actttgctct tgaaggaact ctagctgctg acggcaaaac aacattaaaa 54agaag gcactgttgt tttaagcaaa cacatttcaa actctggaga aataacagtt 6ttaatg actctgacac tactcaggct actaaaaaaa ctggaacatg ggattcaaag 66cactt taacaattag tgtgaatagc cgaaaaacca aaaaccttgt attcacaaaa 72cacaa taacagtaca aaaatacgac tcagcaggca ccaatctaga aggcaaagca 78aatta caacgcttaa agaacttaaa gatgctttaa aata 824 5DNA Borrelia burgdorferi CDS ( atg att atc aat cat aat aca tca gct att aat gct tca aga aat aat 48 Met Ile Ile Asn His Asn Thr Ser Ala Ile Asn Ala Ser Arg Asn Asn att aat gct gct aat ctt agt aaa acc caa gag aag ctt tct agt 96 Ala Ile Asn Ala Ala Asn Leu Ser Lys Thr Gln Glu Lys Leu Ser Ser 2 ggt tac aga att aat cga gct tct gat gat gct gct ggt atg ggg gtt Tyr Arg Ile Asn Arg Ala Ser Asp Asp Ala Ala Gly Met Gly Val 35 4t ggc aag att aat gct caa ata aca ggc tta tca caa gct tct aga Gly Lys Ile Asn Ala Gln Ile Thr Gly Leu Ser Gln Ala Ser Arg 5 aac act tca aaa gct atc aat ttt att cag aca aca gaa gga aat tta 24hr Ser Lys Ala Ile Asn Phe Ile Gln Thr Thr Glu Gly Asn Leu 65 7 aat gaa gta gaa aaa gtt tta gta aga atg aaa gaa tta gca gtt caa 288 Asn Glu Val Glu Lys Val Leu Val Arg Met Lys Glu Leu Ala Val Gln 85 9a ggt aac gga acg tat tca gac gca gac aga ggt tct ata cag att 336 Ser Gly Asn Gly Thr Tyr Ser Asp Ala Asp Arg Gly Ser Ile Gln Ile ata gag caa ctt aca gac gaa att aat aga att gct gat cag gct 384 Glu Ile Glu Gln Leu Thr Asp Glu Ile Asn Arg Ile Ala Asp Gln Ala tat aac caa atg cac atg ttg tca aac aaa tct gct tcc caa aat 432 Gln Tyr Asn Gln Met His Met Leu Ser Asn Lys Ser Ala Ser Gln Asn aaa aca gct gaa gag ctt gga atg cag cct gca aaa att aac aca 48ys Thr Ala Glu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr cca gca tca ctt tca gga tct caa gct tct tgg act tta aga gtt cat 528 Pro Ala Ser Leu Ser Gly Ser Gln Ala Ser Trp Thr Leu Arg Val His gga gca aat caa gat gaa gca att gct gta aat att tat tca gct 576 Val Gly Ala Asn Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ser Ala gtt gca aat ctt ttt gct ggt gag gga gct caa gct gct cag gct 624 Asn Val Ala Asn Leu Phe Ala Gly Glu Gly Ala Gln Ala Ala Gln Ala 2cct gtt caa gag ggt gct caa gaa gaa gga gct cag caa cca aca 672 Ala Pro Val Gln Glu Gly Ala Gln Glu Glu Gly Ala Gln Gln Pro Thr 222ct aca gca cct act caa ggt gga gtt aat tct cct gtt aat gtt 72la Thr Ala Pro Thr Gln Gly Gly Val Asn Ser Pro Val Asn Val 225 234cc aca gtt gat gct aat aca tca ctt gct aaa ata gaa aat gct 768 Thr Thr Thr Val Asp Ala Asn Thr Ser Leu Ala Lys Ile Glu Asn Ala 245 25tt aga atg ata agt gat caa aga gca aat tta ggt gct ttc caa aat 8Arg Met Ile Ser Asp Gln Arg Ala Asn Leu Gly Ala Phe Gln Asn 267tt gaa tct ata aag aat agc act gag tat gct att gaa aat cta 864 Arg Leu Glu Ser Ile Lys Asn Ser Thr Glu Tyr Ala Ile Glu Asn Leu 275 28aa gca tct tat gct caa ata aaa gat gct aca atg aca gat gag gtt 9Ala Ser Tyr Ala Gln Ile Lys Asp Ala Thr Met Thr Asp Glu Val 29gca gct aca act aat agt att tta act caa tct gca atg gca atg 96la Ala Thr Thr Asn Ser Ile Leu Thr Gln Ser Ala Met Ala Met 33att gca cag gct aat caa gtt cct caa tat gtt ttg tca ttg ctt aga le Ala Gln Ala Asn Gln Val Pro Gln Tyr Val Leu Ser Leu Leu Arg

325 33aa 52 336 PRT Borrelia burgdorferi 52 Met Ile Ile Asn His Asn Thr Ser Ala Ile Asn Ala Ser Arg Asn Asn Ile Asn Ala Ala Asn Leu Ser Lys Thr Gln Glu Lys Leu Ser Ser 2 Gly Tyr Arg Ile Asn Arg Ala Ser Asp Asp Ala Ala Gly Met Gly Val 35 4r Gly Lys Ile Asn Ala Gln Ile Thr Gly Leu Ser Gln Ala Ser Arg 5 Asn Thr Ser Lys Ala Ile Asn Phe Ile Gln Thr Thr Glu Gly Asn Leu 65 7 Asn Glu Val Glu Lys Val Leu Val Arg Met Lys Glu Leu Ala Val Gln 85 9r Gly Asn Gly Thr Tyr Ser Asp Ala Asp Arg Gly Ser Ile Gln Ile Ile Glu Gln Leu Thr Asp Glu Ile Asn Arg Ile Ala Asp Gln Ala Tyr Asn Gln Met His Met Leu Ser Asn Lys Ser Ala Ser Gln Asn Lys Thr Ala Glu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr Pro Ala Ser Leu Ser Gly Ser Gln Ala Ser Trp Thr Leu Arg Val His Gly Ala Asn Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ser Ala Val Ala Asn Leu Phe Ala Gly Glu Gly Ala Gln Ala Ala Gln Ala 2Pro Val Gln Glu Gly Ala Gln Glu Glu Gly Ala Gln Gln Pro Thr 222la Thr Ala Pro Thr Gln Gly Gly Val Asn Ser Pro Val Asn Val 225 234hr Thr Val Asp Ala Asn Thr Ser Leu Ala Lys Ile Glu Asn Ala 245 25le Arg Met Ile Ser Asp Gln Arg Ala Asn Leu Gly Ala Phe Gln Asn 267eu Glu Ser Ile Lys Asn Ser Thr Glu Tyr Ala Ile Glu Asn Leu 275 28ys Ala Ser Tyr Ala Gln Ile Lys Asp Ala Thr Met Thr Asp Glu Val 29Ala Ala Thr Thr Asn Ser Ile Leu Thr Gln Ser Ala Met Ala Met 33Ile Ala Gln Ala Asn Gln Val Pro Gln Tyr Val Leu Ser Leu Leu Arg 325 333 A Borrelia burgdorferi CDS ( atg att atc aat cat aat aca tca gct att aat gct tca aga aat aat 48 Met Ile Ile Asn His Asn Thr Ser Ala Ile Asn Ala Ser Arg Asn Asn att aat gct gct aat ctt agt aaa act caa gag aag ctt tct agt 96 Gly Ile Asn Ala Ala Asn Leu Ser Lys Thr Gln Glu Lys Leu Ser Ser 2 ggt tac aga att aat aga gct tct gat gat gct gct ggt atg ggg gtt Tyr Arg Ile Asn Arg Ala Ser Asp Asp Ala Ala Gly Met Gly Val 35 4t ggg aag att aat gct caa ata aga ggt tta tca caa gct tct aga Gly Lys Ile Asn Ala Gln Ile Arg Gly Leu Ser Gln Ala Ser Arg 5 aac act tca aaa gct att aat ttt att cag aca aca gaa gga aat ttg 24hr Ser Lys Ala Ile Asn Phe Ile Gln Thr Thr Glu Gly Asn Leu 65 7 aat gaa gta gaa aaa gtt tta gta aga atg aaa gaa tta gca gtt caa 288 Asn Glu Val Glu Lys Val Leu Val Arg Met Lys Glu Leu Ala Val Gln 85 9a ggt aac ggt aca tat tca gac gca gac aga ggt tct ata caa att 336 Ser Gly Asn Gly Thr Tyr Ser Asp Ala Asp Arg Gly Ser Ile Gln Ile ata gag caa ctt aca gac gaa att aat aga att gct gat caa gct 384 Glu Ile Glu Gln Leu Thr Asp Glu Ile Asn Arg Ile Ala Asp Gln Ala tat aac caa atg cac atg ttg tca aac aaa tct gct tcc caa aat 432 Gln Tyr Asn Gln Met His Met Leu Ser Asn Lys Ser Ala Ser Gln Asn aga aca gct gaa gaa ctt gga atg caa cct gca aaa atc aac aca 48rg Thr Ala Glu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr cca gcg tca ctt tca gga tct caa gct tct tgg act tta aga gtt cat 528 Pro Ala Ser Leu Ser Gly Ser Gln Ala Ser Trp Thr Leu Arg Val His gga gca aat caa gat gaa gcg att gct gta aat att tat gct gct 576 Val Gly Ala Asn Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ala Ala gtt gca aat cta ttc tct ggt gaa gga gct cag gct gct cag act 624 Asn Val Ala Asn Leu Phe Ser Gly Glu Gly Ala Gln Ala Ala Gln Thr 2cct gtt caa gaa ggt gct caa caa gaa gga gct caa caa cca gca 672 Ala Pro Val Gln Glu Gly Ala Gln Gln Glu Gly Ala Gln Gln Pro Ala 222ct aca gcg cct tct cag ggt gga gtt aat tct cct gtt aat gtt 72la Thr Ala Pro Ser Gln Gly Gly Val Asn Ser Pro Val Asn Val 225 234ct aca gtt gac gct aat aca tct ctt gct aaa ata gaa aat gct 768 Thr Thr Thr Val Asp Ala Asn Thr Ser Leu Ala Lys Ile Glu Asn Ala 245 25tt aga atg ata agt gat caa aga gca aat tta ggt gct ttc caa aat 8Arg Met Ile Ser Asp Gln Arg Ala Asn Leu Gly Ala Phe Gln Asn 267tt gag tct ata aag gat agt act gag tat gct att gaa aac cta 864 Arg Leu Glu Ser Ile Lys Asp Ser Thr Glu Tyr Ala Ile Glu Asn Leu 275 28aa gca tct tat gct caa ata aaa gat gct aca atg aca gat gag gtt 9Ala Ser Tyr Ala Gln Ile Lys Asp Ala Thr Met Thr Asp Glu Val 29gca gct aca act aat agt att tta aca caa tgt gca atg gca atg 96la Ala Thr Thr Asn Ser Ile Leu Thr Gln Cys Ala Met Ala Met 33att gcg caa gct aat caa gtt cct caa tat gtt ttg tca ttg ctt aga e Ala Gln Ala Asn Gln Val Pro Gln Tyr Val Leu Ser Leu Leu Arg 325 334 336 PRT Borrelia burgdorferi 54 Met Ile Ile Asn His Asn Thr Ser Ala Ile Asn Ala Ser Arg Asn Asn Ile Asn Ala Ala Asn Leu Ser Lys Thr Gln Glu Lys Leu Ser Ser 2 Gly Tyr Arg Ile Asn Arg Ala Ser Asp Asp Ala Ala Gly Met Gly Val 35 4r Gly Lys Ile Asn Ala Gln Ile Arg Gly Leu Ser Gln Ala Ser Arg 5 Asn Thr Ser Lys Ala Ile Asn Phe Ile Gln Thr Thr Glu Gly Asn Leu 65 7 Asn Glu Val Glu Lys Val Leu Val Arg Met Lys Glu Leu Ala Val Gln 85 9r Gly Asn Gly Thr Tyr Ser Asp Ala Asp Arg Gly Ser Ile Gln Ile Ile Glu Gln Leu Thr Asp Glu Ile Asn Arg Ile Ala Asp Gln Ala Tyr Asn Gln Met His Met Leu Ser Asn Lys Ser Ala Ser Gln Asn Arg Thr Ala Glu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr Pro Ala Ser Leu Ser Gly Ser Gln Ala Ser Trp Thr Leu Arg Val His Gly Ala Asn Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ala Ala Val Ala Asn Leu Phe Ser Gly Glu Gly Ala Gln Ala Ala Gln Thr 2Pro Val Gln Glu Gly Ala Gln Gln Glu Gly Ala Gln Gln Pro Ala 222la Thr Ala Pro Ser Gln Gly Gly Val Asn Ser Pro Val Asn Val 225 234hr Thr Val Asp Ala Asn Thr Ser Leu Ala Lys Ile Glu Asn Ala 245 25le Arg Met Ile Ser Asp Gln Arg Ala Asn Leu Gly Ala Phe Gln Asn 267eu Glu Ser Ile Lys Asp Ser Thr Glu Tyr Ala Ile Glu Asn Leu 275 28ys Ala Ser Tyr Ala Gln Ile Lys Asp Ala Thr Met Thr Asp Glu Val 29Ala Ala Thr Thr Asn Ser Ile Leu Thr Gln Cys Ala Met Ala Met 33Ile Ala Gln Ala Asn Gln Val Pro Gln Tyr Val Leu Ser Leu Leu Arg 325 335 82orrelia burgdorferi 55 atgaaaaaat atttattggg aataggtcta atattagcct taatagcatg caagcaaaat 6cagcc ttgatgaaaa aaacagcgct tcagtagatt tgcctggtga gatgaaagtt gtaagta aagaaaaaga caaagacggt aagtacagtc taaaggcaac agtagacaag gagctaa aaggaacttc tgataaagac aatggttctg gggtgcttga aggtacaaaa 24caaaa gtaaagcaaa attaacaatt gctgacgatc taggtaaaac cacattcgaa 3tcaaag aagatggcaa aacattagtg tcaagaaaag taagttctaa agacaaaaca 36agatg aaatgttcaa tgaaaaaggt gaattgtctg caaaaaccat gacaagagaa 42aacca aacttgaata tacagaaatg aaaagcgatg gaaccggaaa agctaaagaa 48aaaaa actttactct tgaaggaaaa gtagctaatg ataaagtaac attggaagta 54aggaa ccgttacttt aagtaaggaa attgcaaaat ctggagaagt aacagttgct 6atgaca ctaacactac tcaggctact aaaaaaactg gcgcatggga ttcaaaaact 66tttaa caattagtgt taacagcaaa aaaactacac aacttgtgtt tactaaacaa 72aataa ctgtacaaaa atacgactcc gcaggtacca atttagaagg cacagcagtc 78taaaa cacttgatga acttaaaaac gctttaaaat a 82orrelia burgdorferi 56 atgaaaaaat atttattggg aataggtcta atattagcct taatagcatg caagcaaaat 6cagcc ttgatgaaaa aaacagcgct tcagtagatt tgcctggtga gattaaagtt gtaagta aagaaaaaga caaagacggt aagtacagtc taaaggcaac agtagacaag gagctaa aaggaacttc tgataaagac aatggttctg gagtgcttga aggtacaaaa 24caaaa gtaaagcaaa attaacaatt gctgacgatc taagtaaaac cacattcgaa 3tcaaag aagatggcaa aacattagtg tcaagaaaag taagttctaa agacaaaaca 36agatg aaatgttcaa tgaaaaaggt gaattgtctg caaaaaccat gacaagagaa 42aacca aacttgaata tacagaaatg aaaagcgatg gaaccggaaa agctaaagaa 48aaaaa actttactct tgaaggaaaa gtagctaatg ataaagtaac attggaagta 54aggaa ccgttacttt aagtaaggaa attgcaaaat ctggagaagt aacagttgct 6atgaca ctaacactac tcaggctact aaaaaaactg gcgcatggga ttcaaaaact 66tttaa caattagtgt taacagtaaa aaaactacac aacttgtgtt tactaaacaa 72aataa ctgtacaaaa atacgactcc gcaggtacca atttagaagg cacagcagtc 78taaaa cacttgatga acttaaaaac gctttaaaat a 82orrelia burgdorferi 57 atgaaaaaat atttattggg aataggtcta atattagcct taatagcatg caagcaaaat 6cagcc ttgatgaaaa aaacagcgct tcagtagatt tgcctggtga gatgaaagtt gtaagta aagaaaaaga caaagacggt aagtacagtc taaaggcaac agtagacaag gagctaa aaggaacttc tgataaagac aatggttctg gggtgcttga aggtacaaaa 24caaaa gtaaagcaaa attaacaatt gctgacgatc taggtaaaac cacattcgaa 3tcaaag aagatggcaa aacattagtg tcaagaaaag taagttctaa agacaaaaca 36agatg aaatgttcaa tgaaaaaggt gaattgtctg caaaaaccat gacaagagaa 42aacca aacttgaata tacagaaatg aaaagcgatg gaaccggaaa agctaaagaa 48aaaaa actttactct tgaaggaaaa gtagctaatg ataaagtaac attggaagta 54aggaa ccgttacttt aagtaaggaa attgcaaaat ctggagaagt aacagttgct 6atgaca ctaacactac tcaggctact aaaaaaactg gcgcatggga ttcaaaaact 66tttaa caattagtgt taacagcaaa aaaactacac aacttgtgtt tactaaacaa 72aataa ctgtacaaaa atacgactcc gcaggtacca atttagaagg cacagcagtc 78taaaa cacttgatga acttaaaaac gctttaaaat a 822 DNA Borrelia burgdorferi 58 atgaaaaaat atttattggg aataggtcta atattagcct taatagcatg caagcaaaat 6cagcc ttgatgaaaa aaacagcgct tcagtagatt tgcctggtga gatgaaagtt gtaagta aagaaaaaga caaagacggt aagtacagtc taaaggcaac agtagacaag gagctaa aaggaacttc tgataaagac aatggttctg gagtgcttga aggtacaaaa 24caaaa gtaaagcaaa attaacaatt gctgacgatc taagtaaaac cacattcgaa 3tcaaag aagatggcaa aacattagtg tcaagaaaag taagttctaa agacaaaaca 36agatg aaatgttcaa tgaaaaaggt gaattgtctg caaaaaccat gacaagagaa 42aacca aacttgaata tacagaaatg aaaagcgatg gaaccggaaa agctaaagaa 48aaaaa actttactct tgaaggaaaa gtagctaatg ataaagtaac attggaagta 54aggaa ccgttacttt aagtaaggaa attgcaaaat ctggagaagt aacagttgct 6atgaca ctaacactac tcaggctact aaaaaaactg gcgcatggga ttcaaaaact 66tttaa caattagtgt taacagcaaa aaaactacac aacttgtgtt tactaaacaa 72aataa ctgtacaaaa atacgactcc gcaggtacca atttagaagg cacagcagtc 78taaaa cacttgatga acttaaaaac gctttgaaat aa 822 59 37 DNA Artificial Sequence Oligonucleotide Primer 59 ttggatccgg tcaccccatg gctcaatata accaatg 37 6A Artificial Sequence Oligonucleotide Primer 6tccgg tcaccccatg gcttctcaaa atgtaag 37 6A Artificial Sequence Oligonucleotide Primer 6tccgg tgaccaactc cgccttgaga agg 33 62 33 DNA Artificial Sequence Oligonucleotide Primer 62 ttggatccgg tgacctattt gagcataaga tgc 33 63 2rtificial Sequence Oligonucleotide Primer 63 ggtgaattta gttggtaagg 2 DNA Artificial Sequence Oligonucleotide Primer 64 caccagtttc tttaagctgc tcctgc 26 65 2 Borrelia burgdorferi CDS ( atg aaa aaa atg tta cta atc ttt agt ttt ttt ctt att ttc ttg aat 48 Met Lys Lys Met Leu Leu Ile Phe Ser Phe Phe Leu Ile Phe Leu Asn ttt cct gtt agt gca aga gaa gtt gat agg gaa aaa tta aag gac 96 Gly Phe Pro Val Ser Ala Arg Glu Val Asp Arg Glu Lys Leu Lys Asp 2 ttt gtt aat atg gat ctt gag ttt gta aat tat aaa ggc cct tat gat Val Asn Met Asp Leu Glu Phe Val Asn Tyr Lys Gly Pro Tyr Asp 35 4t aca aat aca tat gaa caa ata gtg ggt att ggg gag ttt tta gca Thr Asn Thr Tyr Glu Gln Ile Val Gly Ile Gly Glu Phe Leu Ala 5 aga ccg ttg acc aat tcc aat agc aac tca agt tat tat ggt aaa tat 24ro Leu Thr Asn Ser Asn Ser Asn Ser Ser Tyr Tyr Gly Lys Tyr 65 7 ttt att aat aga ttt att gat gat caa gat aaa aaa gca agc gtt gat 288 Phe Ile Asn Arg Phe Ile Asp Asp Gln Asp Lys Lys Ala Ser Val Asp 85 9t ttt tct att ggt agt aag tca gag ctt gac agt ata ttg aat tta 336 Val Phe Ser Ile Gly Ser Lys Ser Glu Leu Asp Ser Ile Leu Asn Leu aga att ctt aca ggg tat tta ata aag tct ttc gat tat gac agg 384 Arg Arg Ile Leu Thr Gly Tyr Leu Ile Lys Ser Phe Asp Tyr Asp Arg agt gca gaa tta att gct aag gtt att aca ata tat aat gct gtt 432 Ser Ser Ala Glu Leu Ile Ala Lys Val Ile Thr Ile Tyr Asn Ala Val aga gga gat ttg gat tat tat aaa ggg ttt tat att gag gct gct 48rg Gly Asp Leu Asp Tyr Tyr Lys Gly Phe Tyr Ile Glu Ala Ala tta aag tct tta agt aaa gaa aat gca ggt ctt tct agg gtt tat agt 528 Leu Lys Ser Leu Ser Lys Glu Asn Ala Gly Leu Ser Arg Val Tyr Ser tgg gct gga aag aca caa ata ttt att cct ctt aaa aag gat att 576 Gln Trp Ala Gly Lys Thr Gln Ile Phe Ile Pro Leu Lys Lys Asp Ile tct gga aat att gag tct gac att gat att gac agt tta gtt aca 624 Leu Ser Gly Asn Ile Glu Ser Asp Ile Asp Ile Asp Ser Leu Val Thr 2aag gtg gtg gca gct ctt tta agt gaa aat gaa gca ggt gtt aac 672 Asp Lys Val Val Ala Ala Leu Leu Ser Glu Asn Glu Ala Gly Val Asn 222ca aga gat att aca gat att caa ggc gaa act cat aag gca gat 72la Arg Asp Ile Thr Asp Ile Gln Gly Glu Thr His Lys Ala Asp 225 234at aaa att gat att gaa tta gac aat att cat gaa agt gat tcc 768 Gln Asp Lys Ile Asp Ile Glu Leu Asp Asn Ile His Glu Ser Asp Ser 245 25at ata aca gaa act att gaa aat tta agg gat cag ctt gaa aaa gct 8Ile Thr Glu Thr Ile Glu Asn Leu Arg Asp Gln Leu Glu Lys Ala 267at gaa gag cat aaa aaa gag att gaa agt cag gtt gat gct aaa 864 Thr Asp Glu Glu His Lys Lys Glu Ile Glu Ser Gln Val Asp Ala Lys 275 28ag aaa caa aag gaa gag cta gat aaa aag gca ata aat ctt gat aaa 9Lys Gln Lys Glu Glu Leu Asp Lys Lys Ala Ile Asn Leu Asp Lys 29cag caa aaa tta gat tct gct gaa gat aat tta gat gtt caa aga 96ln Gln Lys Leu Asp Ser Ala Glu Asp Asn Leu Asp Val Gln Arg 33aat act gtt aga gag aaa att caa gag gat

att aac gaa att aac aag n Thr Val Arg Glu Lys Ile Gln Glu Asp Ile Asn Glu Ile Asn Lys 325 33aa aag aat tta cca aag cct ggt gat gta agt tct cct aaa gtt gat u Lys Asn Leu Pro Lys Pro Gly Asp Val Ser Ser Pro Lys Val Asp 345aa cta caa ata aaa gag agc ctg gaa gat ttg cag gag cag ctt s Gln Leu Gln Ile Lys Glu Ser Leu Glu Asp Leu Gln Glu Gln Leu 355 36aa gaa act ggt gat gaa aat cag aaa aga gaa att gaa aag caa att s Glu Thr Gly Asp Glu Asn Gln Lys Arg Glu Ile Glu Lys Gln Ile 378tc aaa aaa agt gat gaa aag ctt tta aaa agt aaa gat gat aaa u Ile Lys Lys Ser Asp Glu Lys Leu Leu Lys Ser Lys Asp Asp Lys 385 39agt aaa gat ggt aaa gcc ttg gat ctt gat cga gaa tta aat tct a Ser Lys Asp Gly Lys Ala Leu Asp Leu Asp Arg Glu Leu Asn Ser 44gct tct agc aaa gaa aaa agt aaa gcc aag gaa gaa gaa ata acc s Ala Ser Ser Lys Glu Lys Ser Lys Ala Lys Glu Glu Glu Ile Thr 423gt aag tca cag aaa agc tta ggc gat ttg aat aat gat gaa aat s Gly Lys Ser Gln Lys Ser Leu Gly Asp Leu Asn Asn Asp Glu Asn 435 44tt atg atg cca gaa gat caa aaa tta cct gag gtt aaa aaa tta gat u Met Met Pro Glu Asp Gln Lys Leu Pro Glu Val Lys Lys Leu Asp 456aa aaa gaa ttt aaa cct gtt tct gag gtt gag aaa tta gat aag r Lys Lys Glu Phe Lys Pro Val Ser Glu Val Glu Lys Leu Asp Lys 465 478tc aag tct aat aac aat gtt gga gaa tta tca ccg tta gat aaa e Phe Lys Ser Asn Asn Asn Val Gly Glu Leu Ser Pro Leu Asp Lys 485 49ct tct tat aaa gac att gat tca aaa gag gag aca gtt aat aaa gat r Ser Tyr Lys Asp Ile Asp Ser Lys Glu Glu Thr Val Asn Lys Asp 55aat ttg caa aag act aag cct cag gtt aaa gac caa gtt act tct l Asn Leu Gln Lys Thr Lys Pro Gln Val Lys Asp Gln Val Thr Ser 5525 ttg aat gaa gat ttg act act atg tct ata gat tcc agt agt cct gta u Asn Glu Asp Leu Thr Thr Met Ser Ile Asp Ser Ser Ser Pro Val 534ta gag gtt att gat cca att aca aat tta gga act ctt caa ctt e Leu Glu Val Ile Asp Pro Ile Thr Asn Leu Gly Thr Leu Gln Leu 545 556at tta aat act ggt gtt agg ctt aaa gaa agc act cag caa ggc e Asp Leu Asn Thr Gly Val Arg Leu Lys Glu Ser Thr Gln Gln Gly 565 57tt cag cgg tat gga att tat gaa cgt gaa aaa gat ttg gtt gtt att e Gln Arg Tyr Gly Ile Tyr Glu Arg Glu Lys Asp Leu Val Val Ile 589tg gat tca gga aaa gct aag ctt cag ata ctt gat aaa ctt gaa s Met Asp Ser Gly Lys Ala Lys Leu Gln Ile Leu Asp Lys Leu Glu 595 6aat tta aaa gtg gta tca gag tct aat ttt gag att aat aaa aat tca n Leu Lys Val Val Ser Glu Ser Asn Phe Glu Ile Asn Lys Asn Ser 662tt tat gtt gat tct aaa atg att tta gta gct gtt agg gat aaa r Leu Tyr Val Asp Ser Lys Met Ile Leu Val Ala Val Arg Asp Lys 625 634gt agt aat gat tgg aga ttg gcc aaa ttt tct cct aaa aat tta p Ser Ser Asn Asp Trp Arg Leu Ala Lys Phe Ser Pro Lys Asn Leu 645 65at gag ttt att ctt tca gag aat aaa att atg cct ttt act agc ttt 2 Glu Phe Ile Leu Ser Glu Asn Lys Ile Met Pro Phe Thr Ser Phe 667tg aga aaa aat ttt att tat ttg caa gat gag ttt aaa agt cta 2 Val Arg Lys Asn Phe Ile Tyr Leu Gln Asp Glu Phe Lys Ser Leu 675 68tt att tta gat gta aat act tta aaa aaa gtt aag ta 2 Ile Leu Asp Val Asn Thr Leu Lys Lys Val Lys 69Borrelia burgdorferi 66 Met Lys Lys Met Leu Leu Ile Phe Ser Phe Phe Leu Ile Phe Leu Asn Phe Pro Val Ser Ala Arg Glu Val Asp Arg Glu Lys Leu Lys Asp 2 Phe Val Asn Met Asp Leu Glu Phe Val Asn Tyr Lys Gly Pro Tyr Asp 35 4r Thr Asn Thr Tyr Glu Gln Ile Val Gly Ile Gly Glu Phe Leu Ala 5 Arg Pro Leu Thr Asn Ser Asn Ser Asn Ser Ser Tyr Tyr Gly Lys Tyr 65 7 Phe Ile Asn Arg Phe Ile Asp Asp Gln Asp Lys Lys Ala Ser Val Asp 85 9l Phe Ser Ile Gly Ser Lys Ser Glu Leu Asp Ser Ile Leu Asn Leu Arg Ile Leu Thr Gly Tyr Leu Ile Lys Ser Phe Asp Tyr Asp Arg Ser Ala Glu Leu Ile Ala Lys Val Ile Thr Ile Tyr Asn Ala Val Arg Gly Asp Leu Asp Tyr Tyr Lys Gly Phe Tyr Ile Glu Ala Ala Leu Lys Ser Leu Ser Lys Glu Asn Ala Gly Leu Ser Arg Val Tyr Ser Trp Ala Gly Lys Thr Gln Ile Phe Ile Pro Leu Lys Lys Asp Ile Ser Gly Asn Ile Glu Ser Asp Ile Asp Ile Asp Ser Leu Val Thr 2Lys Val Val Ala Ala Leu Leu Ser Glu Asn Glu Ala Gly Val Asn 222la Arg Asp Ile Thr Asp Ile Gln Gly Glu Thr His Lys Ala Asp 225 234sp Lys Ile Asp Ile Glu Leu Asp Asn Ile His Glu Ser Asp Ser 245 25sn Ile Thr Glu Thr Ile Glu Asn Leu Arg Asp Gln Leu Glu Lys Ala 267sp Glu Glu His Lys Lys Glu Ile Glu Ser Gln Val Asp Ala Lys 275 28ys Lys Gln Lys Glu Glu Leu Asp Lys Lys Ala Ile Asn Leu Asp Lys 29Gln Gln Lys Leu Asp Ser Ala Glu Asp Asn Leu Asp Val Gln Arg 33Asn Thr Val Arg Glu Lys Ile Gln Glu Asp Ile Asn Glu Ile Asn Lys 325 33lu Lys Asn Leu Pro Lys Pro Gly Asp Val Ser Ser Pro Lys Val Asp 345ln Leu Gln Ile Lys Glu Ser Leu Glu Asp Leu Gln Glu Gln Leu 355 36ys Glu Thr Gly Asp Glu Asn Gln Lys Arg Glu Ile Glu Lys Gln Ile 378le Lys Lys Ser Asp Glu Lys Leu Leu Lys Ser Lys Asp Asp Lys 385 39Ser Lys Asp Gly Lys Ala Leu Asp Leu Asp Arg Glu Leu Asn Ser 44Ala Ser Ser Lys Glu Lys Ser Lys Ala Lys Glu Glu Glu Ile Thr 423ly Lys Ser Gln Lys Ser Leu Gly Asp Leu Asn Asn Asp Glu Asn 435 44eu Met Met Pro Glu Asp Gln Lys Leu Pro Glu Val Lys Lys Leu Asp 456ys Lys Glu Phe Lys Pro Val Ser Glu Val Glu Lys Leu Asp Lys 465 478he Lys Ser Asn Asn Asn Val Gly Glu Leu Ser Pro Leu Asp Lys 485 49er Ser Tyr Lys Asp Ile Asp Ser Lys Glu Glu Thr Val Asn Lys Asp 55Asn Leu Gln Lys Thr Lys Pro Gln Val Lys Asp Gln Val Thr Ser 5525 Leu Asn Glu Asp Leu Thr Thr Met Ser Ile Asp Ser Ser Ser Pro Val 534eu Glu Val Ile Asp Pro Ile Thr Asn Leu Gly Thr Leu Gln Leu 545 556sp Leu Asn Thr Gly Val Arg Leu Lys Glu Ser Thr Gln Gln Gly 565 57le Gln Arg Tyr Gly Ile Tyr Glu Arg Glu Lys Asp Leu Val Val Ile 589et Asp Ser Gly Lys Ala Lys Leu Gln Ile Leu Asp Lys Leu Glu 595 6Asn Leu Lys Val Val Ser Glu Ser Asn Phe Glu Ile Asn Lys Asn Ser 662eu Tyr Val Asp Ser Lys Met Ile Leu Val Ala Val Arg Asp Lys 625 634er Ser Asn Asp Trp Arg Leu Ala Lys Phe Ser Pro Lys Asn Leu 645 65sp Glu Phe Ile Leu Ser Glu Asn Lys Ile Met Pro Phe Thr Ser Phe 667al Arg Lys Asn Phe Ile Tyr Leu Gln Asp Glu Phe Lys Ser Leu 675 68al Ile Leu Asp Val Asn Thr Leu Lys Lys Val Lys 69 Borrelia burgdorferi CDS ( atg aaa aaa ttg tta cta atc ttt agt ttt ttt ctt att tct ttg aat 48 Met Lys Lys Leu Leu Leu Ile Phe Ser Phe Phe Leu Ile Ser Leu Asn ttt cct ctt aat tca agg gaa gtt gat aag gaa aaa tta aag gat 96 Gly Phe Pro Leu Asn Ser Arg Glu Val Asp Lys Glu Lys Leu Lys Asp 2 ttt gtt aat atg gat ctt gag ttt gta aac tat aaa ggt cct tat gat Val Asn Met Asp Leu Glu Phe Val Asn Tyr Lys Gly Pro Tyr Asp 35 4t aca aat aca tat gaa caa ata gta ggt att ggt gag ttt tta gca Thr Asn Thr Tyr Glu Gln Ile Val Gly Ile Gly Glu Phe Leu Ala 5 aga cca ttg att aat tcc aat agc aac tca att tat tat ggt aaa tat 24ro Leu Ile Asn Ser Asn Ser Asn Ser Ile Tyr Tyr Gly Lys Tyr 65 7 ttt att aat aga ttt att gat gat caa gat aaa aaa gca agc gtt gat 288 Phe Ile Asn Arg Phe Ile Asp Asp Gln Asp Lys Lys Ala Ser Val Asp 85 9t ttt tct att ggt agt agg tca cag ctt gac agt ata ttg aat cta 336 Val Phe Ser Ile Gly Ser Arg Ser Gln Leu Asp Ser Ile Leu Asn Leu aga att ctt aca ggg tat ttg ata aag tct ttt gat tat gaa aga 384 Arg Arg Ile Leu Thr Gly Tyr Leu Ile Lys Ser Phe Asp Tyr Glu Arg agt gct gaa tta att gct aag gtt att aca ata cat aat gct gtt 432 Ser Ser Ala Glu Leu Ile Ala Lys Val Ile Thr Ile His Asn Ala Val aga ggg gat tta aat tat tat aaa gag gtt tat att gag gct gct 48rg Gly Asp Leu Asn Tyr Tyr Lys Glu Val Tyr Ile Glu Ala Ala tta aag tct tta act aaa gaa aat gca ggt ctt tct aga gtg tac agt 528 Leu Lys Ser Leu Thr Lys Glu Asn Ala Gly Leu Ser Arg Val Tyr Ser tgg gct gga aag aca caa ata ttt att cct ctt aaa aag aat att 576 Gln Trp Ala Gly Lys Thr Gln Ile Phe Ile Pro Leu Lys Lys Asn Ile tct gga aaa gtt gag tct gac att gat att gac agt ttg gtt aca 624 Leu Ser Gly Lys Val Glu Ser Asp Ile Asp Ile Asp Ser Leu Val Thr 2aag gtt gtg gca gct ctt tta agc gag aat gaa gca ggt gtt aac 672 Asp Lys Val Val Ala Ala Leu Leu Ser Glu Asn Glu Ala Gly Val Asn 222ca aga gat att aca gat att caa ggc gaa act cat aaa gca gat 72la Arg Asp Ile Thr Asp Ile Gln Gly Glu Thr His Lys Ala Asp 225 234at aaa att gat att gaa tta gat aat gtt cat aaa agt gat tcc 768 Gln Asp Lys Ile Asp Ile Glu Leu Asp Asn Val His Lys Ser Asp Ser 245 25at ata aca gag act att gag aat tta aga gat cag ctt gaa aag gct 8Ile Thr Glu Thr Ile Glu Asn Leu Arg Asp Gln Leu Glu Lys Ala 267at gaa gag cat aga aaa gag att gaa agt cag gtt gat gct aaa 864 Thr Asp Glu Glu His Arg Lys Glu Ile Glu Ser Gln Val Asp Ala Lys 275 28ag aaa caa aaa gaa gaa cta gat aaa aag gca atc gat ctt gat aaa 9Lys Gln Lys Glu Glu Leu Asp Lys Lys Ala Ile Asp Leu Asp Lys 29caa caa aaa tta gat tct tct gaa gat aat tta gat att caa agg 96ln Gln Lys Leu Asp Ser Ser Glu Asp Asn Leu Asp Ile Gln Arg 33gat act gtt aga gag aag att caa gag gat att gac gag att aat aaa p Thr Val Arg Glu Lys Ile Gln Glu Asp Ile Asp Glu Ile Asn Lys 325 33aa aag aat ttg cca aaa cct ggt gat gta agt tct cct aaa gtt gat u Lys Asn Leu Pro Lys Pro Gly Asp Val Ser Ser Pro Lys Val Asp 345ag cta caa ata aaa gag agt cta gaa gac ttg cag gaa cag ctt s Gln Leu Gln Ile Lys Glu Ser Leu Glu Asp Leu Gln Glu Gln Leu 355 36aa gaa act agc gat gaa aat caa aaa aga gaa att gaa aag caa att s Glu Thr Ser Asp Glu Asn Gln Lys Arg Glu Ile Glu Lys Gln Ile 378tc aaa aaa agt gat gaa gaa ctt tta aaa agt aaa gat cct aaa u Ile Lys Lys Ser Asp Glu Glu Leu Leu Lys Ser Lys Asp Pro Lys 385 39tta gat ctt aat gga gat tta aat tct aaa gtt tct agt aaa gaa a Leu Asp Leu Asn Gly Asp Leu Asn Ser Lys Val Ser Ser Lys Glu 44att aaa ggc aaa gaa gga gaa ata gtc aaa gag gaa tca aag gca s Ile Lys Gly Lys Glu Gly Glu Ile Val Lys Glu Glu Ser Lys Ala 423ta gct gat ttg aat aat gac gaa aat ctt atg agg ccg gaa gat r Leu Ala Asp Leu Asn Asn Asp Glu Asn Leu Met Arg Pro Glu Asp 435 44aa aaa tta tct gag gat aaa aaa tta gat agt aaa aaa aat tta aaa n Lys Leu Ser Glu Asp Lys Lys Leu Asp Ser Lys Lys Asn Leu Lys 456tt tct gag att gag aga gta aat gaa att tcg aag tct aac aac o Val Ser Glu Ile Glu Arg Val Asn Glu Ile Ser Lys Ser Asn Asn 465 478ag att agt gaa tca tca cca tta tat aag cct tct tat agc gat n Glu Ile Ser Glu Ser Ser Pro Leu Tyr Lys Pro Ser Tyr Ser Asp 485 49tg gat tca aaa gag ggt ata gat aat aaa gat gtt aac ttg caa gaa t Asp Ser Lys Glu Gly Ile Asp Asn Lys Asp Val Asn Leu Gln Glu 55aag tct caa act aaa agt caa cct act tct tta aat caa gat ttg r Lys Ser Gln Thr Lys Ser Gln Pro Thr Ser Leu Asn Gln Asp Leu 5525 act act atg tct ata gat tct agt aat cct gta ttt tta gag gtt att r Thr Met Ser Ile Asp Ser Ser Asn Pro Val Phe Leu Glu Val Ile 534ct att aca aat tta gga acg ctt caa ctt att gat ttg aat acc p Pro Ile Thr Asn Leu Gly Thr Leu Gln Leu Ile Asp Leu Asn Thr 545 556tt aga ctt aaa gaa agt act cag caa ggc att cag cgg tat gga y Val Arg Leu Lys Glu Ser Thr Gln Gln Gly Ile Gln Arg Tyr Gly 565 57tt tat gaa cgt gaa aaa gat tta gtt gtt att aaa atg gat tca gga e Tyr Glu Arg Glu Lys Asp Leu Val Val Ile Lys Met Asp Ser Gly 589cc aag ctt caa ata ctt aat aaa ctt gag aat tta aaa gtg ata s Ala Lys Leu Gln Ile Leu Asn Lys Leu Glu Asn Leu Lys Val Ile 595 6tcg gag tct aat ttt gag att aat aaa aat tca tct ctt tat gtt gac r Glu Ser Asn Phe Glu Ile Asn Lys Asn Ser Ser Leu Tyr Val Asp 662aa atg att tta gta gtt gtg aga gat agt ggt aat gtt tgg aga r Lys Met Ile Leu Val Val Val Arg Asp Ser Gly Asn Val Trp Arg 625 634ct aaa ttt tct cct aaa aat tta aat gag ttt att ctt tca gag u Ala Lys Phe Ser Pro Lys Asn Leu Asn Glu Phe Ile Leu Ser Glu 645 65at aaa att ttg cct ttt act agc ttt tct gtg aga aag aat ttt att 2 Lys Ile Leu Pro Phe Thr Ser Phe Ser Val Arg Lys Asn Phe Ile 667tg cag gat gag ttt aaa agt ctt att act tta gat gta aat act 2 Leu Gln Asp Glu Phe Lys Ser Leu Ile Thr Leu Asp Val Asn Thr 675 68ta aaa aaa gtt aag ta 2 Lys Lys Val Lys 693 PRT Borrelia burgdorferi 68 Met Lys Lys Leu Leu

Leu Ile Phe Ser Phe Phe Leu Ile Ser Leu Asn Phe Pro Leu Asn Ser Arg Glu Val Asp Lys Glu Lys Leu Lys Asp 2 Phe Val Asn Met Asp Leu Glu Phe Val Asn Tyr Lys Gly Pro Tyr Asp 35 4r Thr Asn Thr Tyr Glu Gln Ile Val Gly Ile Gly Glu Phe Leu Ala 5 Arg Pro Leu Ile Asn Ser Asn Ser Asn Ser Ile Tyr Tyr Gly Lys Tyr 65 7 Phe Ile Asn Arg Phe Ile Asp Asp Gln Asp Lys Lys Ala Ser Val Asp 85 9l Phe Ser Ile Gly Ser Arg Ser Gln Leu Asp Ser Ile Leu Asn Leu Arg Ile Leu Thr Gly Tyr Leu Ile Lys Ser Phe Asp Tyr Glu Arg Ser Ala Glu Leu Ile Ala Lys Val Ile Thr Ile His Asn Ala Val Arg Gly Asp Leu Asn Tyr Tyr Lys Glu Val Tyr Ile Glu Ala Ala Leu Lys Ser Leu Thr Lys Glu Asn Ala Gly Leu Ser Arg Val Tyr Ser Trp Ala Gly Lys Thr Gln Ile Phe Ile Pro Leu Lys Lys Asn Ile Ser Gly Lys Val Glu Ser Asp Ile Asp Ile Asp Ser Leu Val Thr 2Lys Val Val Ala Ala Leu Leu Ser Glu Asn Glu Ala Gly Val Asn 222la Arg Asp Ile Thr Asp Ile Gln Gly Glu Thr His Lys Ala Asp 225 234sp Lys Ile Asp Ile Glu Leu Asp Asn Val His Lys Ser Asp Ser 245 25sn Ile Thr Glu Thr Ile Glu Asn Leu Arg Asp Gln Leu Glu Lys Ala 267sp Glu Glu His Arg Lys Glu Ile Glu Ser Gln Val Asp Ala Lys 275 28ys Lys Gln Lys Glu Glu Leu Asp Lys Lys Ala Ile Asp Leu Asp Lys 29Gln Gln Lys Leu Asp Ser Ser Glu Asp Asn Leu Asp Ile Gln Arg 33Asp Thr Val Arg Glu Lys Ile Gln Glu Asp Ile Asp Glu Ile Asn Lys 325 33lu Lys Asn Leu Pro Lys Pro Gly Asp Val Ser Ser Pro Lys Val Asp 345ln Leu Gln Ile Lys Glu Ser Leu Glu Asp Leu Gln Glu Gln Leu 355 36ys Glu Thr Ser Asp Glu Asn Gln Lys Arg Glu Ile Glu Lys Gln Ile 378le Lys Lys Ser Asp Glu Glu Leu Leu Lys Ser Lys Asp Pro Lys 385 39Leu Asp Leu Asn Gly Asp Leu Asn Ser Lys Val Ser Ser Lys Glu 44Ile Lys Gly Lys Glu Gly Glu Ile Val Lys Glu Glu Ser Lys Ala 423eu Ala Asp Leu Asn Asn Asp Glu Asn Leu Met Arg Pro Glu Asp 435 44ln Lys Leu Ser Glu Asp Lys Lys Leu Asp Ser Lys Lys Asn Leu Lys 456al Ser Glu Ile Glu Arg Val Asn Glu Ile Ser Lys Ser Asn Asn 465 478lu Ile Ser Glu Ser Ser Pro Leu Tyr Lys Pro Ser Tyr Ser Asp 485 49et Asp Ser Lys Glu Gly Ile Asp Asn Lys Asp Val Asn Leu Gln Glu 55Lys Ser Gln Thr Lys Ser Gln Pro Thr Ser Leu Asn Gln Asp Leu 5525 Thr Thr Met Ser Ile Asp Ser Ser Asn Pro Val Phe Leu Glu Val Ile 534ro Ile Thr Asn Leu Gly Thr Leu Gln Leu Ile Asp Leu Asn Thr 545 556al Arg Leu Lys Glu Ser Thr Gln Gln Gly Ile Gln Arg Tyr Gly 565 57le Tyr Glu Arg Glu Lys Asp Leu Val Val Ile Lys Met Asp Ser Gly 589la Lys Leu Gln Ile Leu Asn Lys Leu Glu Asn Leu Lys Val Ile 595 6Ser Glu Ser Asn Phe Glu Ile Asn Lys Asn Ser Ser Leu Tyr Val Asp 662ys Met Ile Leu Val Val Val Arg Asp Ser Gly Asn Val Trp Arg 625 634la Lys Phe Ser Pro Lys Asn Leu Asn Glu Phe Ile Leu Ser Glu 645 65sn Lys Ile Leu Pro Phe Thr Ser Phe Ser Val Arg Lys Asn Phe Ile 667eu Gln Asp Glu Phe Lys Ser Leu Ile Thr Leu Asp Val Asn Thr 675 68eu Lys Lys Val Lys 699orrelia burgdorferi CDS (989) 69 atg aaa aaa atg tta cta atc ttt agt ttt ttt ctt gtt ttt tta aat 48 Met Lys Lys Met Leu Leu Ile Phe Ser Phe Phe Leu Val Phe Leu Asn ttt cct ctt aat gca agg gaa gtt gat aag gaa aaa tta aag gac 96 Gly Phe Pro Leu Asn Ala Arg Glu Val Asp Lys Glu Lys Leu Lys Asp 2 ttt gtt aat atg gat ctt gaa ttt gtt aat tac aag ggt cct tat gat Val Asn Met Asp Leu Glu Phe Val Asn Tyr Lys Gly Pro Tyr Asp 35 4t aca gat aca tat gaa caa ata gta ggt att ggg gag ttt tta gca Thr Asp Thr Tyr Glu Gln Ile Val Gly Ile Gly Glu Phe Leu Ala 5 agg ccg ttg aac aat tcc aat agt aat tca agt tat tat ggt aaa tat 24ro Leu Asn Asn Ser Asn Ser Asn Ser Ser Tyr Tyr Gly Lys Tyr 65 7 ttt gtt aat aga ttt att gac gat caa gat aaa aaa gca agt gtt gat 288 Phe Val Asn Arg Phe Ile Asp Asp Gln Asp Lys Lys Ala Ser Val Asp 85 9t ttt tct att ggt agt aag tca gag ctt gat agt ata tta aat cta 336 Ile Phe Ser Ile Gly Ser Lys Ser Glu Leu Asp Ser Ile Leu Asn Leu aga att ctt aca ggg tat tta atg aag tct ttt gat tat gag agg 384 Arg Arg Ile Leu Thr Gly Tyr Leu Met Lys Ser Phe Asp Tyr Glu Arg agt gcg gaa tta att gct aaa gct att aca ata tat aat gct gtt 432 Ser Ser Ala Glu Leu Ile Ala Lys Ala Ile Thr Ile Tyr Asn Ala Val aga gga gat tta gat tat tac aaa gag ttt tat att gag gct tct 48rg Gly Asp Leu Asp Tyr Tyr Lys Glu Phe Tyr Ile Glu Ala Ser ttg aag tct ttg act aaa gaa aat gca ggt ctt tct agg gtg tac agt 528 Leu Lys Ser Leu Thr Lys Glu Asn Ala Gly Leu Ser Arg Val Tyr Ser tgg gct ggg aag aca caa ata ttt att cct ctt aaa aag aat att 576 Gln Trp Ala Gly Lys Thr Gln Ile Phe Ile Pro Leu Lys Lys Asn Ile tct gga aat gtt gag tct gac att gat att gat agt ttg gtt aca 624 Leu Ser Gly Asn Val Glu Ser Asp Ile Asp Ile Asp Ser Leu Val Thr 2aag gtg gtg gca gct ctt tta agt gag aat gaa tca ggt gtt aac 672 Asp Lys Val Val Ala Ala Leu Leu Ser Glu Asn Glu Ser Gly Val Asn 222ca aga gat att aca gac att caa ggc gaa act cat aaa gca gat 72la Arg Asp Ile Thr Asp Ile Gln Gly Glu Thr His Lys Ala Asp 225 234at aaa att gat att gaa tta gat aat ttt cat gaa agt gat tcc 768 Gln Asp Lys Ile Asp Ile Glu Leu Asp Asn Phe His Glu Ser Asp Ser 245 25at ata aca gaa act att gag aat tta agg gat cag ctt gaa aaa gct 8Ile Thr Glu Thr Ile Glu Asn Leu Arg Asp Gln Leu Glu Lys Ala 267at gaa gag cat aaa aaa gag att gaa agt cag gtt gat gct aaa 864 Thr Asp Glu Glu His Lys Lys Glu Ile Glu Ser Gln Val Asp Ala Lys 275 28ag aaa caa aag gaa gaa tta gat aaa aag gca att gat ctt gat aaa 9Lys Gln Lys Glu Glu Leu Asp Lys Lys Ala Ile Asp Leu Asp Lys 29caa caa aaa tta gat ttt gct gaa gat aat cta gat att caa agg 96ln Gln Lys Leu Asp Phe Ala Glu Asp Asn Leu Asp Ile Gln Arg 33gat act gtt aga gag aag ctt caa gaa aat att aac gag act aat aag p Thr Val Arg Glu Lys Leu Gln Glu Asn Ile Asn Glu Thr Asn Lys 325 33aa aag aat tta cca aag cct ggt gat gta agt tct cct aag gtt gat u Lys Asn Leu Pro Lys Pro Gly Asp Val Ser Ser Pro Lys Val Asp 345ag ttg cag ata aaa gag agt cta gaa gat ttg caa gag cag ctt s Gln Leu Gln Ile Lys Glu Ser Leu Glu Asp Leu Gln Glu Gln Leu 355 36aa gaa gct agt gat gaa aat caa aaa aga gaa ata gaa aag caa att s Glu Ala Ser Asp Glu Asn Gln Lys Arg Glu Ile Glu Lys Gln Ile 378tc aaa aaa aat gat gaa gaa ctt ttt aaa aat aaa gat cat aaa u Ile Lys Lys Asn Asp Glu Glu Leu Phe Lys Asn Lys Asp His Lys 385 39tta gat ctt aag caa gaa tta aat tct aaa gct tct agt aaa gaa a Leu Asp Leu Lys Gln Glu Leu Asn Ser Lys Ala Ser Ser Lys Glu 44att gaa ggc gaa gaa gag gat aaa gaa tta gat agt aaa aaa aat s Ile Glu Gly Glu Glu Glu Asp Lys Glu Leu Asp Ser Lys Lys Asn 423ag cct gtt tct gag gct gat aaa gta gat aaa att tcc aag tct u Glu Pro Val Ser Glu Ala Asp Lys Val Asp Lys Ile Ser Lys Ser 435 44ac aac aat gag gtt agt aaa tta tcc ccg tta gat gag cct tct tat n Asn Asn Glu Val Ser Lys Leu Ser Pro Leu Asp Glu Pro Ser Tyr 456ac att gat tcg aaa gag ggt gta gat aac aaa gat gtt gat ttg r Asp Ile Asp Ser Lys Glu Gly Val Asp Asn Lys Asp Val Asp Leu 465 478aa act aaa ccc caa gtt gaa agt caa cct act tcg tta aat gaa n Lys Thr Lys Pro Gln Val Glu Ser Gln Pro Thr Ser Leu Asn Glu 485 49at ttg att gat gtg tct ata gat tcc agt aat cct gtc ttt tta gag p Leu Ile Asp Val Ser Ile Asp Ser Ser Asn Pro Val Phe Leu Glu 55atc gat ccg att aca aat tta gga acg ctt caa ctt att gat ttg l Ile Asp Pro Ile Thr Asn Leu Gly Thr Leu Gln Leu Ile Asp Leu 5525 aat acc ggt gtt aga ctt aaa gaa agt gct caa caa ggt att cag cga n Thr Gly Val Arg Leu Lys Glu Ser Ala Gln Gln Gly Ile Gln Arg 534ga att tat gaa cgt gaa aaa gat ttg gtt gtt att aaa ata gat r Gly Ile Tyr Glu Arg Glu Lys Asp Leu Val Val Ile Lys Ile Asp 545 556ga aaa gct aag ctt cag ata ctt gat aaa ctc gag aat tta aaa r Gly Lys Ala Lys Leu Gln Ile Leu Asp Lys Leu Glu Asn Leu Lys 565 57tg ata tca gag tct aat ttt gag att aat aaa aat tca tct ctt tat l Ile Ser Glu Ser Asn Phe Glu Ile Asn Lys Asn Ser Ser Leu Tyr 589ac tct aga atg att tta gta gtt gtt aag gac gat agt aat gct l Asp Ser Arg Met Ile Leu Val Val Val Lys Asp Asp Ser Asn Ala 595 6tgg aga ttg gct aaa ttt tct cct aaa aat tta gat gaa ttt att ctg p Arg Leu Ala Lys Phe Ser Pro Lys Asn Leu Asp Glu Phe Ile Leu 662aa aat aaa att ttg cct ttt act agc ttt gct gtg aga aag aat r Glu Asn Lys Ile Leu Pro Phe Thr Ser Phe Ala Val Arg Lys Asn 625 634tt tat ttg caa gat gaa ctt aaa agc tta gtt act tta gat gta e Ile Tyr Leu Gln Asp Glu Leu Lys Ser Leu Val Thr Leu Asp Val 645 65at act tta aaa aaa gtt aag ta n Thr Leu Lys Lys Val Lys 663 PRT Borrelia burgdorferi 7ys Lys Met Leu Leu Ile Phe Ser Phe Phe Leu Val Phe Leu Asn Phe Pro Leu Asn Ala Arg Glu Val Asp Lys Glu Lys Leu Lys Asp 2 Phe Val Asn Met Asp Leu Glu Phe Val Asn Tyr Lys Gly Pro Tyr Asp 35 4r Thr Asp Thr Tyr Glu Gln Ile Val Gly Ile Gly Glu Phe Leu Ala 5 Arg Pro Leu Asn Asn Ser Asn Ser Asn Ser Ser Tyr Tyr Gly Lys Tyr 65 7 Phe Val Asn Arg Phe Ile Asp Asp Gln Asp Lys Lys Ala Ser Val Asp 85 9e Phe Ser Ile Gly Ser Lys Ser Glu Leu Asp Ser Ile Leu Asn Leu Arg Ile Leu Thr Gly Tyr Leu Met Lys Ser Phe Asp Tyr Glu Arg Ser Ala Glu Leu Ile Ala Lys Ala Ile Thr Ile Tyr Asn Ala Val Arg Gly Asp Leu Asp Tyr Tyr Lys Glu Phe Tyr Ile Glu Ala Ser Leu Lys Ser Leu Thr Lys Glu Asn Ala Gly Leu Ser Arg Val Tyr Ser Trp Ala Gly Lys Thr Gln Ile Phe Ile Pro Leu Lys Lys Asn Ile Ser Gly Asn Val Glu Ser Asp Ile Asp Ile Asp Ser Leu Val Thr 2Lys Val Val Ala Ala Leu Leu Ser Glu Asn Glu Ser Gly Val Asn 222la Arg Asp Ile Thr Asp Ile Gln Gly Glu Thr His Lys Ala Asp 225 234sp Lys Ile Asp Ile Glu Leu Asp Asn Phe His Glu Ser Asp Ser 245 25sn Ile Thr Glu Thr Ile Glu Asn Leu Arg Asp Gln Leu Glu Lys Ala 267sp Glu Glu His Lys Lys Glu Ile Glu Ser Gln Val Asp Ala Lys 275 28ys Lys Gln Lys Glu Glu Leu Asp Lys Lys Ala Ile Asp Leu Asp Lys 29Gln Gln Lys Leu Asp Phe Ala Glu Asp Asn Leu Asp Ile Gln Arg 33Asp Thr Val Arg Glu Lys Leu Gln Glu Asn Ile Asn Glu Thr Asn Lys 325 33lu Lys Asn Leu Pro Lys Pro Gly Asp Val Ser Ser Pro Lys Val Asp 345ln Leu Gln Ile Lys Glu Ser Leu Glu Asp Leu Gln Glu Gln Leu 355 36ys Glu Ala Ser Asp Glu Asn Gln Lys Arg Glu Ile Glu Lys Gln Ile 378le Lys Lys Asn Asp Glu Glu Leu Phe Lys Asn Lys Asp His Lys 385 39Leu Asp Leu Lys Gln Glu Leu Asn Ser Lys Ala Ser Ser Lys Glu 44Ile Glu Gly Glu Glu Glu Asp Lys Glu Leu Asp Ser Lys Lys Asn 423lu Pro Val Ser Glu Ala Asp Lys Val Asp Lys Ile Ser Lys Ser 435 44sn Asn Asn Glu Val Ser Lys Leu Ser Pro Leu Asp Glu Pro Ser Tyr 456sp Ile Asp Ser Lys Glu Gly Val Asp Asn Lys Asp Val Asp Leu 465 478ys Thr Lys Pro Gln Val Glu Ser Gln Pro Thr Ser Leu Asn Glu 485 49sp Leu Ile Asp Val Ser Ile Asp Ser Ser Asn Pro Val Phe Leu Glu 55Ile Asp Pro Ile Thr Asn Leu Gly Thr Leu Gln Leu Ile Asp Leu 5525 Asn Thr Gly Val Arg Leu Lys Glu Ser Ala Gln Gln Gly Ile Gln Arg 534ly Ile Tyr Glu Arg Glu Lys Asp Leu Val Val Ile Lys Ile Asp 545 556ly Lys Ala Lys Leu Gln Ile Leu Asp Lys Leu Glu Asn Leu Lys 565 57al Ile Ser Glu Ser Asn Phe Glu Ile Asn Lys Asn Ser Ser Leu Tyr 589sp Ser Arg Met Ile Leu Val Val Val Lys Asp Asp Ser Asn Ala 595 6Trp Arg Leu Ala Lys Phe Ser Pro Lys Asn Leu Asp Glu Phe Ile Leu 662lu Asn Lys Ile Leu Pro Phe Thr Ser Phe Ala Val Arg Lys Asn 625 634le Tyr Leu Gln Asp Glu Leu Lys Ser Leu Val Thr Leu Asp Val 645 65sn Thr Leu Lys Lys Val Lys 668orrelia burgdorferi CDS ( atg aaa aaa atg tta cta atc ttt agt ttt ttt ctt att

tct ttg aat 48 Met Lys Lys Met Leu Leu Ile Phe Ser Phe Phe Leu Ile Ser Leu Asn ttt ccc ctt aat gca agg gaa gtt gat aag gaa aaa tta aag gac 96 Gly Phe Pro Leu Asn Ala Arg Glu Val Asp Lys Glu Lys Leu Lys Asp 2 ttt gtt aat atg gat ctt gag ttt gta aac tat aaa ggt cct tat gat Val Asn Met Asp Leu Glu Phe Val Asn Tyr Lys Gly Pro Tyr Asp 35 4t aca aat aca tat gaa caa ata gta ggt att ggt gag ttt tta gca Thr Asn Thr Tyr Glu Gln Ile Val Gly Ile Gly Glu Phe Leu Ala 5 aga cca ttg att aat ttc aat agc aac tca agt tat tat ggt aaa tat 24ro Leu Ile Asn Phe Asn Ser Asn Ser Ser Tyr Tyr Gly Lys Tyr 65 7 ttt att aat aga ttt att gac gat caa gat aaa aaa gca agc gtt gat 288 Phe Ile Asn Arg Phe Ile Asp Asp Gln Asp Lys Lys Ala Ser Val Asp 85 9t ttt tct att agt agt aag tca cag ctt gac agt ata ttg aat tta 336 Val Phe Ser Ile Ser Ser Lys Ser Gln Leu Asp Ser Ile Leu Asn Leu aga att ctt aca ggg tat ttg ata aag tct ttt gat tat gaa aga 384 Arg Arg Ile Leu Thr Gly Tyr Leu Ile Lys Ser Phe Asp Tyr Glu Arg agt gct gaa tta att gcc aag gtt att aca ata cat aat gct gtt 432 Ser Ser Ala Glu Leu Ile Ala Lys Val Ile Thr Ile His Asn Ala Val aga ggt gat tta aat tat tat aaa gag ttt tat att gag tct gct 48rg Gly Asp Leu Asn Tyr Tyr Lys Glu Phe Tyr Ile Glu Ser Ala tta aag tct tta act aaa gaa aat gca ggt ctt tct aga gtg tac agt 528 Leu Lys Ser Leu Thr Lys Glu Asn Ala Gly Leu Ser Arg Val Tyr Ser tgg gct gga aag aca caa ata ttt att cct ctt aaa aag aat att 576 Gln Trp Ala Gly Lys Thr Gln Ile Phe Ile Pro Leu Lys Lys Asn Ile tct gga aaa att gag tct gac att gat att gat agt ttg gtt aca 624 Leu Ser Gly Lys Ile Glu Ser Asp Ile Asp Ile Asp Ser Leu Val Thr 2aag gtt gtg gca gct ctt tta agc gaa aat gaa gca ggt gtt aac 672 Asp Lys Val Val Ala Ala Leu Leu Ser Glu Asn Glu Ala Gly Val Asn 222ca agg gat att aca gat att caa gga gaa act cat aaa gca gat 72la Arg Asp Ile Thr Asp Ile Gln Gly Glu Thr His Lys Ala Asp 225 234at aaa att gat att gaa tta gat aat gtt cat gaa agt gat tcc 768 Gln Asp Lys Ile Asp Ile Glu Leu Asp Asn Val His Glu Ser Asp Ser 245 25at ata aca gaa act att gag aat tta aga gat cag ctt gaa aag gct 8Ile Thr Glu Thr Ile Glu Asn Leu Arg Asp Gln Leu Glu Lys Ala 267at gaa gag cat aga aaa gag att gaa agt caa gtt gat gct aaa 864 Thr Asp Glu Glu His Arg Lys Glu Ile Glu Ser Gln Val Asp Ala Lys 275 28ag aaa caa aaa gaa gaa cta gat aaa aag gca atc gat ctt gat aaa 9Lys Gln Lys Glu Glu Leu Asp Lys Lys Ala Ile Asp Leu Asp Lys 29caa caa aaa tta gat ttt tct gaa gat aat tta gat att caa agg 96ln Gln Lys Leu Asp Phe Ser Glu Asp Asn Leu Asp Ile Gln Arg 33gat act gtt aga gag aag att caa gag gat att aac gag att aat aag sp Thr Val Arg Glu Lys Ile Gln Glu Asp Ile Asn Glu Ile Asn Lys 325 33aa aag aat tta cca aaa cct ggt gat gta agt tct cct aaa gtt gat lu Lys Asn Leu Pro Lys Pro Gly Asp Val Ser Ser Pro Lys Val Asp 345ag cta caa ata aaa gag agt cta gaa gac ttg cag gag cag ctt ys Gln Leu Gln Ile Lys Glu Ser Leu Glu Asp Leu Gln Glu Gln Leu 355 36aa gaa act agc gat gaa aat caa aaa aga gaa att gaa aag caa att ys Glu Thr Ser Asp Glu Asn Gln Lys Arg Glu Ile Glu Lys Gln Ile 378tc aaa aaa agt gat gaa gaa ctt tta aaa agc aaa gat cct aaa lu Ile Lys Lys Ser Asp Glu Glu Leu Leu Lys Ser Lys Asp Pro Lys 385 39tta gat ctt aat cga gat tta aat tct aaa gct tct agt aaa gaa la Leu Asp Leu Asn Arg Asp Leu Asn Ser Lys Ala Ser Ser Lys Glu 44att aaa ggc aaa gaa aaa gaa ata gtc aaa gag aaa tca aag gta ys Ile Lys Gly Lys Glu Lys Glu Ile Val Lys Glu Lys Ser Lys Val 423ta ggt gat ttg gat aat gac gaa acc ctt atg acg ccg gaa gat er Leu Gly Asp Leu Asp Asn Asp Glu Thr Leu Met Thr Pro Glu Asp 435 44aa aaa tta tct gag gat aaa aaa tta gat agt aaa aaa aat tta aaa ln Lys Leu Ser Glu Asp Lys Lys Leu Asp Ser Lys Lys Asn Leu Lys 456tt tct gag att gag aga gta aat gaa att tca aag tct aac aac ro Val Ser Glu Ile Glu Arg Val Asn Glu Ile Ser Lys Ser Asn Asn 465 478ag gtt agc aaa tca tca cca tta gat aag cct tct tat agt gat sn Glu Val Ser Lys Ser Ser Pro Leu Asp Lys Pro Ser Tyr Ser Asp 485 49tc gat tca aaa gag gtt gta gat aat aaa gat gtt aat ttg caa gaa le Asp Ser Lys Glu Val Val Asp Asn Lys Asp Val Asn Leu Gln Glu 55aag cct caa gct aaa agt caa tct act tct tta aat caa gat ttg hr Lys Pro Gln Ala Lys Ser Gln Ser Thr Ser Leu Asn Gln Asp Leu 5525 att act atg tct ata gat tct agt aat cct gta ttt tta gag gtt att le Thr Met Ser Ile Asp Ser Ser Asn Pro Val Phe Leu Glu Val Ile 534ct att aca aat tta gga atg ctt caa ctt att gat tta aat act sp Pro Ile Thr Asn Leu Gly Met Leu Gln Leu Ile Asp Leu Asn Thr 545 556tt aga ctt aaa gaa agc act cag caa ggc att cag cgt tat gga ly Val Arg Leu Lys Glu Ser Thr Gln Gln Gly Ile Gln Arg Tyr Gly 565 57tt tat gaa cgt gaa aaa gat tta gtt gtt att aaa atg gat tca gga le Tyr Glu Arg Glu Lys Asp Leu Val Val Ile Lys Met Asp Ser Gly 589ct aag ctt caa ata ctt aat aaa ctt gag aat tta aaa gtg ata ys Ala Lys Leu Gln Ile Leu Asn Lys Leu Glu Asn Leu Lys Val Ile 595 6tca gag tct aat ttt gag att aat aaa aat tca tct ctt tat gtt gac er Glu Ser Asn Phe Glu Ile Asn Lys Asn Ser Ser Leu Tyr Val Asp 662aa atg att tta gta gct gtg aaa gat agt ggt aat gtt tgg aga er Lys Met Ile Leu Val Ala Val Lys Asp Ser Gly Asn Val Trp Arg 625 634ct aaa ttt tct cct aaa aat tta gat gag ttt att ctt tca gag eu Ala Lys Phe Ser Pro Lys Asn Leu Asp Glu Phe Ile Leu Ser Glu 645 65at aaa att ttg cct ttt act agc ttt tct gtg aga aag aat ttt att 2 Lys Ile Leu Pro Phe Thr Ser Phe Ser Val Arg Lys Asn Phe Ile 667tg caa gat gag ttt aaa agt ctt att act tta gat gta aat act 2 Leu Gln Asp Glu Phe Lys Ser Leu Ile Thr Leu Asp Val Asn Thr 675 68ta aaa aaa gtt aag ta 2 Lys Lys Val Lys 693 PRT Borrelia burgdorferi 72 Met Lys Lys Met Leu Leu Ile Phe Ser Phe Phe Leu Ile Ser Leu Asn Phe Pro Leu Asn Ala Arg Glu Val Asp Lys Glu Lys Leu Lys Asp 2 Phe Val Asn Met Asp Leu Glu Phe Val Asn Tyr Lys Gly Pro Tyr Asp 35 4r Thr Asn Thr Tyr Glu Gln Ile Val Gly Ile Gly Glu Phe Leu Ala 5 Arg Pro Leu Ile Asn Phe Asn Ser Asn Ser Ser Tyr Tyr Gly Lys Tyr 65 7 Phe Ile Asn Arg Phe Ile Asp Asp Gln Asp Lys Lys Ala Ser Val Asp 85 9l Phe Ser Ile Ser Ser Lys Ser Gln Leu Asp Ser Ile Leu Asn Leu Arg Ile Leu Thr Gly Tyr Leu Ile Lys Ser Phe Asp Tyr Glu Arg Ser Ala Glu Leu Ile Ala Lys Val Ile Thr Ile His Asn Ala Val Arg Gly Asp Leu Asn Tyr Tyr Lys Glu Phe Tyr Ile Glu Ser Ala Leu Lys Ser Leu Thr Lys Glu Asn Ala Gly Leu Ser Arg Val Tyr Ser Trp Ala Gly Lys Thr Gln Ile Phe Ile Pro Leu Lys Lys Asn Ile Ser Gly Lys Ile Glu Ser Asp Ile Asp Ile Asp Ser Leu Val Thr 2Lys Val Val Ala Ala Leu Leu Ser Glu Asn Glu Ala Gly Val Asn 222la Arg Asp Ile Thr Asp Ile Gln Gly Glu Thr His Lys Ala Asp 225 234sp Lys Ile Asp Ile Glu Leu Asp Asn Val His Glu Ser Asp Ser 245 25sn Ile Thr Glu Thr Ile Glu Asn Leu Arg Asp Gln Leu Glu Lys Ala 267sp Glu Glu His Arg Lys Glu Ile Glu Ser Gln Val Asp Ala Lys 275 28ys Lys Gln Lys Glu Glu Leu Asp Lys Lys Ala Ile Asp Leu Asp Lys 29Gln Gln Lys Leu Asp Phe Ser Glu Asp Asn Leu Asp Ile Gln Arg 33Asp Thr Val Arg Glu Lys Ile Gln Glu Asp Ile Asn Glu Ile Asn Lys 325 33lu Lys Asn Leu Pro Lys Pro Gly Asp Val Ser Ser Pro Lys Val Asp 345ln Leu Gln Ile Lys Glu Ser Leu Glu Asp Leu Gln Glu Gln Leu 355 36ys Glu Thr Ser Asp Glu Asn Gln Lys Arg Glu Ile Glu Lys Gln Ile 378le Lys Lys Ser Asp Glu Glu Leu Leu Lys Ser Lys Asp Pro Lys 385 39Leu Asp Leu Asn Arg Asp Leu Asn Ser Lys Ala Ser Ser Lys Glu 44Ile Lys Gly Lys Glu Lys Glu Ile Val Lys Glu Lys Ser Lys Val 423eu Gly Asp Leu Asp Asn Asp Glu Thr Leu Met Thr Pro Glu Asp 435 44ln Lys Leu Ser Glu Asp Lys Lys Leu Asp Ser Lys Lys Asn Leu Lys 456al Ser Glu Ile Glu Arg Val Asn Glu Ile Ser Lys Ser Asn Asn 465 478lu Val Ser Lys Ser Ser Pro Leu Asp Lys Pro Ser Tyr Ser Asp 485 49le Asp Ser Lys Glu Val Val Asp Asn Lys Asp Val Asn Leu Gln Glu 55Lys Pro Gln Ala Lys Ser Gln Ser Thr Ser Leu Asn Gln Asp Leu 5525 Ile Thr Met Ser Ile Asp Ser Ser Asn Pro Val Phe Leu Glu Val Ile 534ro Ile Thr Asn Leu Gly Met Leu Gln Leu Ile Asp Leu Asn Thr 545 556al Arg Leu Lys Glu Ser Thr Gln Gln Gly Ile Gln Arg Tyr Gly 565 57le Tyr Glu Arg Glu Lys Asp Leu Val Val Ile Lys Met Asp Ser Gly 589la Lys Leu Gln Ile Leu Asn Lys Leu Glu Asn Leu Lys Val Ile 595 6Ser Glu Ser Asn Phe Glu Ile Asn Lys Asn Ser Ser Leu Tyr Val Asp 662ys Met Ile Leu Val Ala Val Lys Asp Ser Gly Asn Val Trp Arg 625 634la Lys Phe Ser Pro Lys Asn Leu Asp Glu Phe Ile Leu Ser Glu 645 65sn Lys Ile Leu Pro Phe Thr Ser Phe Ser Val Arg Lys Asn Phe Ile 667eu Gln Asp Glu Phe Lys Ser Leu Ile Thr Leu Asp Val Asn Thr 675 68eu Lys Lys Val Lys 69Borrelia burgdorferi CDS ( atg aaa aaa atg tta cta atc ttt agt ttt ttt ctt gtt ttt tta aat 48 Met Lys Lys Met Leu Leu Ile Phe Ser Phe Phe Leu Val Phe Leu Asn ttt cct ctt aat gca agg gaa gtt gat aag gaa aaa tta aag gac 96 Gly Phe Pro Leu Asn Ala Arg Glu Val Asp Lys Glu Lys Leu Lys Asp 2 ttt gtt aat atg gat ctt gaa ttt gtt aat tac aag ggt cct tat gat Val Asn Met Asp Leu Glu Phe Val Asn Tyr Lys Gly Pro Tyr Asp 35 4t aca aat aca tat gaa caa ata gta ggt att ggg gag ttt tta gca Thr Asn Thr Tyr Glu Gln Ile Val Gly Ile Gly Glu Phe Leu Ala 5 agg ccg ttg atc aat tcc aat agt aat tca agt tat tat ggt aaa tat 24ro Leu Ile Asn Ser Asn Ser Asn Ser Ser Tyr Tyr Gly Lys Tyr 65 7 ttt gtt aat aga ttt att gac gat caa gat aaa aaa gca agt gtt gat 288 Phe Val Asn Arg Phe Ile Asp Asp Gln Asp Lys Lys Ala Ser Val Asp 85 9t ttt tct att ggt agt aag tca gag ctt gat agt ata tta aat cta 336 Ile Phe Ser Ile Gly Ser Lys Ser Glu Leu Asp Ser Ile Leu Asn Leu aga att ctt aca ggg tat tta atg aag tct ttt gat tat gag agg 384 Arg Arg Ile Leu Thr Gly Tyr Leu Met Lys Ser Phe Asp Tyr Glu Arg agt gcg gaa tta att gct aaa gct att aca ata tat aat gct gtt 432 Ser Ser Ala Glu Leu Ile Ala Lys Ala Ile Thr Ile Tyr Asn Ala Val aga gga gat tta gat tat tac aaa gag ttt tat att gag gct tct 48rg Gly Asp Leu Asp Tyr Tyr Lys Glu Phe Tyr Ile Glu Ala Ser ttg aag tct ttg act aaa gaa aat gca ggt ctt tct agg gtg tac agt 528 Leu Lys Ser Leu Thr Lys Glu Asn Ala Gly Leu Ser Arg Val Tyr Ser tgg gct ggg aag aca caa ata ttt att cct ctt aaa aag aat att 576 Gln Trp Ala Gly Lys Thr Gln Ile Phe Ile Pro Leu Lys Lys Asn Ile tct gga aat gtt gag tct gac att gat att gat agt ttg gtt aca 624 Leu Ser Gly Asn Val Glu Ser Asp Ile Asp Ile Asp Ser Leu Val Thr 2aag gtg gtg gca gct ctt tta agt gag aat gaa tca ggt gtt aac 672 Asp Lys Val Val Ala Ala Leu Leu Ser Glu Asn Glu Ser Gly Val Asn 222ca aga gat att aca gac att caa ggc gaa act cat aaa gca gat 72la Arg Asp Ile Thr Asp Ile Gln Gly Glu Thr His Lys Ala Asp 225 234at aaa att gat att gaa tta gat aat att cat gaa agt gat tcc 768 Gln Asp Lys Ile Asp Ile Glu Leu Asp Asn Ile His Glu Ser Asp Ser 245 25at ata aca gaa act att gag aat tta agg gat cag ctt gaa aaa gct 8Ile Thr Glu Thr Ile Glu Asn Leu Arg Asp Gln Leu Glu Lys Ala 267at gaa gag cat aaa aaa gag att gaa agt cag gtt gat gct aaa 864 Thr Asp Glu Glu His Lys Lys Glu Ile Glu Ser Gln Val Asp Ala Lys 275 28ag aaa caa aag gaa gaa tta gat aaa aag gca att gat ctt gat aaa 9Lys Gln Lys Glu Glu Leu Asp Lys Lys Ala Ile Asp Leu Asp Lys 29caa caa aaa tta gat ttt gct gaa gat aat cta gat att caa agg 96ln Gln Lys Leu Asp Phe Ala Glu Asp Asn Leu Asp Ile Gln Arg 33gat act gtt aga gag aag ctt caa gag aat att aac gag act aat aag p Thr Val Arg Glu Lys Leu Gln Glu Asn Ile Asn Glu Thr Asn Lys 325 33aa aag aat tta cca aag cct ggt gat gta agt tct cct aaa gtt gat u Lys Asn Leu Pro Lys Pro Gly Asp Val Ser Ser Pro Lys Val Asp 345aa cta caa ata aaa gag agc ctg gaa gat ttg cag gag cag ctt s Gln Leu Gln Ile Lys Glu Ser Leu Glu Asp Leu Gln Glu Gln Leu 355 36aa gaa act ggt gat gaa aat cag aaa aga gaa att gaa aag caa att s Glu Thr Gly Asp Glu Asn Gln Lys Arg Glu Ile Glu Lys Gln Ile 378tc aaa aaa agt gat gaa aag ctt tta aaa agt aaa gat gat aaa u Ile Lys Lys Ser

Asp Glu Lys Leu Leu Lys Ser Lys Asp Asp Lys 385 39agt aaa gat ggt aaa gcc ttg gat ctt gat cga gaa tta aat tct a Ser Lys Asp Gly Lys Ala Leu Asp Leu Asp Arg Glu Leu Asn Ser 44gct tct agc aaa gaa aaa agt aaa gcc aag gaa gaa gaa ata acc s Ala Ser Ser Lys Glu Lys Ser Lys Ala Lys Glu Glu Glu Ile Thr 423gt aag tca cag aaa agc tta ggc gat ttg aat aat gat gaa aat s Gly Lys Ser Gln Lys Ser Leu Gly Asp Leu Asn Asn Asp Glu Asn 435 44tt atg atg cca gaa gat caa aaa tta cct gag gtt aaa aaa tta gat u Met Met Pro Glu Asp Gln Lys Leu Pro Glu Val Lys Lys Leu Asp 456aa aaa gaa ttt aaa cct gtt tct gag gtt gag aaa tta gat aag r Lys Lys Glu Phe Lys Pro Val Ser Glu Val Glu Lys Leu Asp Lys 465 478tc aag tct aat aac aat gtt gga gaa tta tca ccg tta gat aaa e Phe Lys Ser Asn Asn Asn Val Gly Glu Leu Ser Pro Leu Asp Lys 485 49ct tct tat aaa gac att gat tca aaa gag gag aca gtt aat aaa gat r Ser Tyr Lys Asp Ile Asp Ser Lys Glu Glu Thr Val Asn Lys Asp 55aat ttg caa aag act aag cct cag gtt aaa gac caa gtt act tct l Asn Leu Gln Lys Thr Lys Pro Gln Val Lys Asp Gln Val Thr Ser 5525 ttg aat gaa gat ttg act act atg tct ata gat tcc agt agt cct gta u Asn Glu Asp Leu Thr Thr Met Ser Ile Asp Ser Ser Ser Pro Val 534ta gag gtt att gat cca att aca aat tta gga act ctt caa ctt e Leu Glu Val Ile Asp Pro Ile Thr Asn Leu Gly Thr Leu Gln Leu 545 556at tta aat act ggt gtt agg ctt aaa gaa agc act cag caa ggc e Asp Leu Asn Thr Gly Val Arg Leu Lys Glu Ser Thr Gln Gln Gly 565 57tt cag cgg tat gga att tat gaa cgt gaa aaa gat ttg gtt gtt att e Gln Arg Tyr Gly Ile Tyr Glu Arg Glu Lys Asp Leu Val Val Ile 589tg gat tca gga aaa gct aag ctt cag ata ctt gat aaa ctt gaa s Met Asp Ser Gly Lys Ala Lys Leu Gln Ile Leu Asp Lys Leu Glu 595 6aat tta aaa gtg gta tca gag tct aat ttt gag att aat aaa aat tca n Leu Lys Val Val Ser Glu Ser Asn Phe Glu Ile Asn Lys Asn Ser 662tt tat gtt gat tct aaa atg att tta gta gct gtt agg gat aaa r Leu Tyr Val Asp Ser Lys Met Ile Leu Val Ala Val Arg Asp Lys 625 634gt agt aat gat tgg aga ttg gcc aaa ttt tct cct aaa aat tta p Ser Ser Asn Asp Trp Arg Leu Ala Lys Phe Ser Pro Lys Asn Leu 645 65at gag ttt att ctt tca gag aat aaa att atg cct ttt act agc ttt 2 Glu Phe Ile Leu Ser Glu Asn Lys Ile Met Pro Phe Thr Ser Phe 667tg aga aaa aat ttt att tat ttg caa gat gag ttt aaa agt cta 2 Val Arg Lys Asn Phe Ile Tyr Leu Gln Asp Glu Phe Lys Ser Leu 675 68tt att tta gat gta aat act tta aaa aaa gtt aag taaagcc 2 Ile Leu Asp Val Asn Thr Leu Lys Lys Val Lys 69Borrelia burgdorferi 74 Met Lys Lys Met Leu Leu Ile Phe Ser Phe Phe Leu Val Phe Leu Asn Phe Pro Leu Asn Ala Arg Glu Val Asp Lys Glu Lys Leu Lys Asp 2 Phe Val Asn Met Asp Leu Glu Phe Val Asn Tyr Lys Gly Pro Tyr Asp 35 4r Thr Asn Thr Tyr Glu Gln Ile Val Gly Ile Gly Glu Phe Leu Ala 5 Arg Pro Leu Ile Asn Ser Asn Ser Asn Ser Ser Tyr Tyr Gly Lys Tyr 65 7 Phe Val Asn Arg Phe Ile Asp Asp Gln Asp Lys Lys Ala Ser Val Asp 85 9e Phe Ser Ile Gly Ser Lys Ser Glu Leu Asp Ser Ile Leu Asn Leu Arg Ile Leu Thr Gly Tyr Leu Met Lys Ser Phe Asp Tyr Glu Arg Ser Ala Glu Leu Ile Ala Lys Ala Ile Thr Ile Tyr Asn Ala Val Arg Gly Asp Leu Asp Tyr Tyr Lys Glu Phe Tyr Ile Glu Ala Ser Leu Lys Ser Leu Thr Lys Glu Asn Ala Gly Leu Ser Arg Val Tyr Ser Trp Ala Gly Lys Thr Gln Ile Phe Ile Pro Leu Lys Lys Asn Ile Ser Gly Asn Val Glu Ser Asp Ile Asp Ile Asp Ser Leu Val Thr 2Lys Val Val Ala Ala Leu Leu Ser Glu Asn Glu Ser Gly Val Asn 222la Arg Asp Ile Thr Asp Ile Gln Gly Glu Thr His Lys Ala Asp 225 234sp Lys Ile Asp Ile Glu Leu Asp Asn Ile His Glu Ser Asp Ser 245 25sn Ile Thr Glu Thr Ile Glu Asn Leu Arg Asp Gln Leu Glu Lys Ala 267sp Glu Glu His Lys Lys Glu Ile Glu Ser Gln Val Asp Ala Lys 275 28ys Lys Gln Lys Glu Glu Leu Asp Lys Lys Ala Ile Asp Leu Asp Lys 29Gln Gln Lys Leu Asp Phe Ala Glu Asp Asn Leu Asp Ile Gln Arg 33Asp Thr Val Arg Glu Lys Leu Gln Glu Asn Ile Asn Glu Thr Asn Lys 325 33lu Lys Asn Leu Pro Lys Pro Gly Asp Val Ser Ser Pro Lys Val Asp 345ln Leu Gln Ile Lys Glu Ser Leu Glu Asp Leu Gln Glu Gln Leu 355 36ys Glu Thr Gly Asp Glu Asn Gln Lys Arg Glu Ile Glu Lys Gln Ile 378le Lys Lys Ser Asp Glu Lys Leu Leu Lys Ser Lys Asp Asp Lys 385 39Ser Lys Asp Gly Lys Ala Leu Asp Leu Asp Arg Glu Leu Asn Ser 44Ala Ser Ser Lys Glu Lys Ser Lys Ala Lys Glu Glu Glu Ile Thr 423ly Lys Ser Gln Lys Ser Leu Gly Asp Leu Asn Asn Asp Glu Asn 435 44eu Met Met Pro Glu Asp Gln Lys Leu Pro Glu Val Lys Lys Leu Asp 456ys Lys Glu Phe Lys Pro Val Ser Glu Val Glu Lys Leu Asp Lys 465 478he Lys Ser Asn Asn Asn Val Gly Glu Leu Ser Pro Leu Asp Lys 485 49er Ser Tyr Lys Asp Ile Asp Ser Lys Glu Glu Thr Val Asn Lys Asp 55Asn Leu Gln Lys Thr Lys Pro Gln Val Lys Asp Gln Val Thr Ser 5525 Leu Asn Glu Asp Leu Thr Thr Met Ser Ile Asp Ser Ser Ser Pro Val 534eu Glu Val Ile Asp Pro Ile Thr Asn Leu Gly Thr Leu Gln Leu 545 556sp Leu Asn Thr Gly Val Arg Leu Lys Glu Ser Thr Gln Gln Gly 565 57le Gln Arg Tyr Gly Ile Tyr Glu Arg Glu Lys Asp Leu Val Val Ile 589et Asp Ser Gly Lys Ala Lys Leu Gln Ile Leu Asp Lys Leu Glu 595 6Asn Leu Lys Val Val Ser Glu Ser Asn Phe Glu Ile Asn Lys Asn Ser 662eu Tyr Val Asp Ser Lys Met Ile Leu Val Ala Val Arg Asp Lys 625 634er Ser Asn Asp Trp Arg Leu Ala Lys Phe Ser Pro Lys Asn Leu 645 65sp Glu Phe Ile Leu Ser Glu Asn Lys Ile Met Pro Phe Thr Ser Phe 667al Arg Lys Asn Phe Ile Tyr Leu Gln Asp Glu Phe Lys Ser Leu 675 68al Ile Leu Asp Val Asn Thr Leu Lys Lys Val Lys 69 Borrelia burgdorferi CDS ( atg aaa aaa atg tta cta atc ttt agt ttt ttt ctt att ttt ttg aat 48 Met Lys Lys Met Leu Leu Ile Phe Ser Phe Phe Leu Ile Phe Leu Asn ttt cct ctt aat gca agg aaa gtt gat aag gaa aaa tta aag gat 96 Gly Phe Pro Leu Asn Ala Arg Lys Val Asp Lys Glu Lys Leu Lys Asp 2 ttt gtt aat atg gat ctt gag ttt gta aat tat aaa ggt cct tat gat Val Asn Met Asp Leu Glu Phe Val Asn Tyr Lys Gly Pro Tyr Asp 35 4t aca aat acg tat gaa caa ata gtg ggt att ggg gag ttt tta gca Thr Asn Thr Tyr Glu Gln Ile Val Gly Ile Gly Glu Phe Leu Ala 5 aga ccg ctg acc aat tcc aat agc aac tca agt tat tat ggc aaa tat 24ro Leu Thr Asn Ser Asn Ser Asn Ser Ser Tyr Tyr Gly Lys Tyr 65 7 ttt att aat aga ttt att gat gat caa gat aaa aaa gca agt gtt gat 288 Phe Ile Asn Arg Phe Ile Asp Asp Gln Asp Lys Lys Ala Ser Val Asp 85 9t ttt tct ata agc agc aaa tca gag ctt gac agt ata ttg aat tta 336 Val Phe Ser Ile Ser Ser Lys Ser Glu Leu Asp Ser Ile Leu Asn Leu aga att ctt aca ggg tat ata ata aag tct ttc gat tat gac agg 384 Arg Arg Ile Leu Thr Gly Tyr Ile Ile Lys Ser Phe Asp Tyr Asp Arg agt gca gaa tta att gct aag gtt att aca ata tat aat gct gtt 432 Ser Ser Ala Glu Leu Ile Ala Lys Val Ile Thr Ile Tyr Asn Ala Val aga gga gat ttg gat tat tat aaa ggg ttt tat att gag cct gct 48rg Gly Asp Leu Asp Tyr Tyr Lys Gly Phe Tyr Ile Glu Pro Ala ttg aag tct tta act aaa gaa aac gca ggt ctt tct agg gtt tac agt 528 Leu Lys Ser Leu Thr Lys Glu Asn Ala Gly Leu Ser Arg Val Tyr Ser tgg gct gga aag act caa ata ttt att cct ctt aaa aag gat att 576 Gln Trp Ala Gly Lys Thr Gln Ile Phe Ile Pro Leu Lys Lys Asp Ile tct gga aat att gaa tct gac att gat att gac agt ttg gtt aca 624 Leu Ser Gly Asn Ile Glu Ser Asp Ile Asp Ile Asp Ser Leu Val Thr 2aag gtg ata gca gct ctt tta agc gaa aat gaa gca ggc gtt aac 672 Asp Lys Val Ile Ala Ala Leu Leu Ser Glu Asn Glu Ala Gly Val Asn 222ca aga gat att aca gat att caa ggc gaa act cat aag gca gat 72la Arg Asp Ile Thr Asp Ile Gln Gly Glu Thr His Lys Ala Asp 225 234at aag att gat act gaa tta gac aat atc cat gaa agc gat tct 768 Gln Asp Lys Ile Asp Thr Glu Leu Asp Asn Ile His Glu Ser Asp Ser 245 25at ata aca gaa act att gaa aat tta agg gat cag ctt gaa aaa gct 8Ile Thr Glu Thr Ile Glu Asn Leu Arg Asp Gln Leu Glu Lys Ala 267at gaa gag cat aaa aaa gag att gaa agt cag gtt gat gct aaa 864 Thr Asp Glu Glu His Lys Lys Glu Ile Glu Ser Gln Val Asp Ala Lys 275 28ag aaa gaa aag gaa gag cta gat aaa aag gca atc aat ctt gat aaa 9Lys Glu Lys Glu Glu Leu Asp Lys Lys Ala Ile Asn Leu Asp Lys 29cag caa aaa tta gac tct gct gaa gat aat tta gat gtt caa aga 96ln Gln Lys Leu Asp Ser Ala Glu Asp Asn Leu Asp Val Gln Arg 33gat act gtt aga gag aaa att caa gag gat att aat gag att aat aag p Thr Val Arg Glu Lys Ile Gln Glu Asp Ile Asn Glu Ile Asn Lys 325 33aa aag aat ttg cca aaa cct ggt gat gta agt tct cct aaa gtt gat u Lys Asn Leu Pro Lys Pro Gly Asp Val Ser Ser Pro Lys Val Asp 345aa ctg caa ata aaa gag agt cta gaa gat ttg cag gag cag ctt s Gln Leu Gln Ile Lys Glu Ser Leu Glu Asp Leu Gln Glu Gln Leu 355 36aa gaa gct ggt gat gaa aat cag aaa aga gaa att gag aag caa att s Glu Ala Gly Asp Glu Asn Gln Lys Arg Glu Ile Glu Lys Gln Ile 378tc aaa aaa agg gac gaa gaa ctt tta aaa agt aaa gat ggc aaa u Ile Lys Lys Arg Asp Glu Glu Leu Leu Lys Ser Lys Asp Gly Lys 385 39agt aaa gat tat gaa gca tta gat ctt gat cga gaa tta tcc aaa l Ser Lys Asp Tyr Glu Ala Leu Asp Leu Asp Arg Glu Leu Ser Lys 44tct agt aaa gaa aaa agt aag gtc aag gaa gaa gaa ata act aaa a Ser Ser Lys Glu Lys Ser Lys Val Lys Glu Glu Glu Ile Thr Lys 423aa tca cgg gca agc tta ggc gat ttg aat aat gat aaa aac ctt y Lys Ser Arg Ala Ser Leu Gly Asp Leu Asn Asn Asp Lys Asn Leu 435 44tg ttg cca gaa gat caa aaa tta cct gaa gat aaa aaa ttg gat agt t Leu Pro Glu Asp Gln Lys Leu Pro Glu Asp Lys Lys Leu Asp Ser 456ta gat ggt aaa aaa gaa ttt aaa cca gtt tct gag gtt gaa aaa s Leu Asp Gly Lys Lys Glu Phe Lys Pro Val Ser Glu Val Glu Lys 465 478at aag att tcc aag tct aat aac aat gag gtt ggc aag tta tca u Asp Lys Ile Ser Lys Ser Asn Asn Asn Glu Val Gly Lys Leu Ser 485 49ca tta gat aag cct tct tat gat gat att gat tca aaa gag gag gta o Leu Asp Lys Pro Ser Tyr Asp Asp Ile Asp Ser Lys Glu Glu Val 55aat aaa gct att aat ttg caa aag atc gac cct aaa gtt aaa gac p Asn Lys Ala Ile Asn Leu Gln Lys Ile Asp Pro Lys Val Lys Asp 5525 caa act act tct ttg aat gaa gat ttg gat aaa gat ttg act act atg n Thr Thr Ser Leu Asn Glu Asp Leu Asp Lys Asp Leu Thr Thr Met 534ta gat tcc agc agt cct gta ttt cta gag gtt att gat cct att r Ile Asp Ser Ser Ser Pro Val Phe Leu Glu Val Ile Asp Pro Ile 545 556at tta gga acc ctg cag ctt att gat tta aat act ggg gtt agg r Asn Leu Gly Thr Leu Gln Leu Ile Asp Leu Asn Thr Gly Val Arg 565 57tt aag gaa agc act cag caa ggc att cag cgg tat gga att tat gaa u Lys Glu Ser Thr Gln Gln Gly Ile Gln Arg Tyr Gly Ile Tyr Glu 589aa aaa gat ttg gtt gtt att aaa atg gat tca gga aag gct aag g Glu Lys Asp Leu Val Val Ile Lys Met Asp Ser Gly Lys Ala Lys 595 6ctt caa ata ctt aat aag ctt gaa aat ttg aaa gtg gta tca gag tct u Gln Ile Leu Asn Lys Leu Glu Asn Leu Lys Val Val Ser Glu Ser 662tt gag atc aat aaa aat tca tct ctt tat gtt gac tct aaa atg n Phe Glu Ile Asn Lys Asn Ser Ser Leu Tyr Val Asp Ser Lys Met 625 634tg gca gct gtt aga gat aag gat gat agc aat gct tgg aga ttg e Leu Ala Ala Val Arg Asp Lys Asp Asp Ser Asn Ala Trp Arg Leu 645 65ct aaa ttt tct cct aaa aat ttg gat gag ttt att ctt tca gag aat 2 Lys Phe Ser Pro Lys Asn Leu Asp Glu Phe Ile Leu Ser Glu Asn 667tt ttg cct ttt act agc ttt tct gtg aga aaa aat ttt att tat 2 Ile Leu Pro Phe Thr Ser Phe Ser Val Arg Lys Asn Phe Ile Tyr 675 68tg caa gat gag ctt aaa aat cta gtt att tta gat gta aat act tta 2 Gln Asp Glu Leu Lys Asn Leu Val Ile Leu Asp Val Asn Thr Leu 69aaa gtt aag ta 2 Lys Val Lys 7Borrelia burgdorferi 76 Met Lys Lys Met Leu Leu Ile Phe Ser Phe Phe Leu Ile Phe Leu Asn Phe Pro Leu Asn Ala Arg Lys Val Asp Lys Glu Lys Leu Lys Asp 2 Phe Val Asn Met Asp Leu Glu Phe Val Asn Tyr Lys Gly Pro Tyr Asp 35 4r Thr Asn Thr Tyr Glu Gln Ile Val Gly Ile Gly Glu Phe Leu Ala 5 Arg Pro Leu Thr Asn Ser Asn Ser Asn Ser Ser Tyr Tyr Gly Lys Tyr 65 7 Phe Ile Asn Arg Phe Ile Asp Asp Gln Asp

Lys Lys Ala Ser Val Asp 85 9l Phe Ser Ile Ser Ser Lys Ser Glu Leu Asp Ser Ile Leu Asn Leu Arg Ile Leu Thr Gly Tyr Ile Ile Lys Ser Phe Asp Tyr Asp Arg Ser Ala Glu Leu Ile Ala Lys Val Ile Thr Ile Tyr Asn Ala Val Arg Gly Asp Leu Asp Tyr Tyr Lys Gly Phe Tyr Ile Glu Pro Ala Leu Lys Ser Leu Thr Lys Glu Asn Ala Gly Leu Ser Arg Val Tyr Ser Trp Ala Gly Lys Thr Gln Ile Phe Ile Pro Leu Lys Lys Asp Ile Ser Gly Asn Ile Glu Ser Asp Ile Asp Ile Asp Ser Leu Val Thr 2Lys Val Ile Ala Ala Leu Leu Ser Glu Asn Glu Ala Gly Val Asn 222la Arg Asp Ile Thr Asp Ile Gln Gly Glu Thr His Lys Ala Asp 225 234sp Lys Ile Asp Thr Glu Leu Asp Asn Ile His Glu Ser Asp Ser 245 25sn Ile Thr Glu Thr Ile Glu Asn Leu Arg Asp Gln Leu Glu Lys Ala 267sp Glu Glu His Lys Lys Glu Ile Glu Ser Gln Val Asp Ala Lys 275 28ys Lys Glu Lys Glu Glu Leu Asp Lys Lys Ala Ile Asn Leu Asp Lys 29Gln Gln Lys Leu Asp Ser Ala Glu Asp Asn Leu Asp Val Gln Arg 33Asp Thr Val Arg Glu Lys Ile Gln Glu Asp Ile Asn Glu Ile Asn Lys 325 33lu Lys Asn Leu Pro Lys Pro Gly Asp Val Ser Ser Pro Lys Val Asp 345ln Leu Gln Ile Lys Glu Ser Leu Glu Asp Leu Gln Glu Gln Leu 355 36ys Glu Ala Gly Asp Glu Asn Gln Lys Arg Glu Ile Glu Lys Gln Ile 378le Lys Lys Arg Asp Glu Glu Leu Leu Lys Ser Lys Asp Gly Lys 385 39Ser Lys Asp Tyr Glu Ala Leu Asp Leu Asp Arg Glu Leu Ser Lys 44Ser Ser Lys Glu Lys Ser Lys Val Lys Glu Glu Glu Ile Thr Lys 423ys Ser Arg Ala Ser Leu Gly Asp Leu Asn Asn Asp Lys Asn Leu 435 44et Leu Pro Glu Asp Gln Lys Leu Pro Glu Asp Lys Lys Leu Asp Ser 456eu Asp Gly Lys Lys Glu Phe Lys Pro Val Ser Glu Val Glu Lys 465 478sp Lys Ile Ser Lys Ser Asn Asn Asn Glu Val Gly Lys Leu Ser 485 49ro Leu Asp Lys Pro Ser Tyr Asp Asp Ile Asp Ser Lys Glu Glu Val 55Asn Lys Ala Ile Asn Leu Gln Lys Ile Asp Pro Lys Val Lys Asp 5525 Gln Thr Thr Ser Leu Asn Glu Asp Leu Asp Lys Asp Leu Thr Thr Met 534le Asp Ser Ser Ser Pro Val Phe Leu Glu Val Ile Asp Pro Ile 545 556sn Leu Gly Thr Leu Gln Leu Ile Asp Leu Asn Thr Gly Val Arg 565 57eu Lys Glu Ser Thr Gln Gln Gly Ile Gln Arg Tyr Gly Ile Tyr Glu 589lu Lys Asp Leu Val Val Ile Lys Met Asp Ser Gly Lys Ala Lys 595 6Leu Gln Ile Leu Asn Lys Leu Glu Asn Leu Lys Val Val Ser Glu Ser 662he Glu Ile Asn Lys Asn Ser Ser Leu Tyr Val Asp Ser Lys Met 625 634eu Ala Ala Val Arg Asp Lys Asp Asp Ser Asn Ala Trp Arg Leu 645 65la Lys Phe Ser Pro Lys Asn Leu Asp Glu Phe Ile Leu Ser Glu Asn 667le Leu Pro Phe Thr Ser Phe Ser Val Arg Lys Asn Phe Ile Tyr 675 68eu Gln Asp Glu Leu Lys Asn Leu Val Ile Leu Asp Val Asn Thr Leu 69Lys Val Lys 799orrelia burgdorferi CDS (989) 77 atg aaa aaa atg tta cta atc ttt agt ttt ttt ctt gtt ttt tta aat 48 Met Lys Lys Met Leu Leu Ile Phe Ser Phe Phe Leu Val Phe Leu Asn ttt cct ctt aat gca agg gaa gtt gat aag gaa aaa tta aag gac 96 Gly Phe Pro Leu Asn Ala Arg Glu Val Asp Lys Glu Lys Leu Lys Asp 2 ttt gtt aat atg gat ctt gaa ttt gtt aat tac aag ggt cct tat gat Val Asn Met Asp Leu Glu Phe Val Asn Tyr Lys Gly Pro Tyr Asp 35 4t aca aat aca tat gaa caa ata gta ggt att ggg gag ttt tta gca Thr Asn Thr Tyr Glu Gln Ile Val Gly Ile Gly Glu Phe Leu Ala 5 agg ccg ttg atc aat tcc aat agt aat tca agt tat tat ggt aaa tat 24ro Leu Ile Asn Ser Asn Ser Asn Ser Ser Tyr Tyr Gly Lys Tyr 65 7 ttt gtt aat aga ttt att gac gat caa gat aaa aaa gca agt gtt gat 288 Phe Val Asn Arg Phe Ile Asp Asp Gln Asp Lys Lys Ala Ser Val Asp 85 9t ttt tct att ggt agt aag tca gag ctt gat agt ata tta aat cta 336 Ile Phe Ser Ile Gly Ser Lys Ser Glu Leu Asp Ser Ile Leu Asn Leu aga att ctt aca ggg tat tta atg aag tct ttt gat tat gag agg 384 Arg Arg Ile Leu Thr Gly Tyr Leu Met Lys Ser Phe Asp Tyr Glu Arg agt gcg gaa tta att gct aaa gct att aca ata tat aat gct gtt 432 Ser Ser Ala Glu Leu Ile Ala Lys Ala Ile Thr Ile Tyr Asn Ala Val aga gga gat tta gat tat tac aaa gag ttt tat att gag gct tct 48rg Gly Asp Leu Asp Tyr Tyr Lys Glu Phe Tyr Ile Glu Ala Ser ttg aag tct ttg act aaa gaa aat gca ggt ctt tct agg gtg tac agt 528 Leu Lys Ser Leu Thr Lys Glu Asn Ala Gly Leu Ser Arg Val Tyr Ser tgg gct ggg aag aca caa ata ttt att cct ctt aaa aag aat att 576 Gln Trp Ala Gly Lys Thr Gln Ile Phe Ile Pro Leu Lys Lys Asn Ile tct gga aat gtt gag tct gac att gat att gat agt ttg gtt aca 624 Leu Ser Gly Asn Val Glu Ser Asp Ile Asp Ile Asp Ser Leu Val Thr 2aag gtg gtg gca gct ctt tta agt gag aat gaa tca ggt gtt aac 672 Asp Lys Val Val Ala Ala Leu Leu Ser Glu Asn Glu Ser Gly Val Asn 222ca aga gat att aca gac att caa ggc gaa act cat aaa gca gat 72la Arg Asp Ile Thr Asp Ile Gln Gly Glu Thr His Lys Ala Asp 225 234at aaa att gat att gaa tta gat aat ttt cat gaa agt gat tcc 768 Gln Asp Lys Ile Asp Ile Glu Leu Asp Asn Phe His Glu Ser Asp Ser 245 25at ata aca gaa act att gag aat tta agg gat cag ctt gaa aaa gct 8Ile Thr Glu Thr Ile Glu Asn Leu Arg Asp Gln Leu Glu Lys Ala 267at gaa gag cat aaa aaa gag att gaa agt cag gtt gat gct aaa 864 Thr Asp Glu Glu His Lys Lys Glu Ile Glu Ser Gln Val Asp Ala Lys 275 28ag aaa caa aag gaa gaa tta gat aaa aag gca att gat ctt gat aaa 9Lys Gln Lys Glu Glu Leu Asp Lys Lys Ala Ile Asp Leu Asp Lys 29caa caa aaa tta gat ttt gct gaa gat aat cta gat att caa agg 96ln Gln Lys Leu Asp Phe Ala Glu Asp Asn Leu Asp Ile Gln Arg 33gat act gtt aga gag aag ctt caa gaa aat att aac gag act aat aag p Thr Val Arg Glu Lys Leu Gln Glu Asn Ile Asn Glu Thr Asn Lys 325 33aa aag aat tta cca aag cct ggt gat gta agt tct cct aag gtt gat u Lys Asn Leu Pro Lys Pro Gly Asp Val Ser Ser Pro Lys Val Asp 345ag ttg cag ata aaa gag agt cta gaa gat ttg caa gag cag ctt s Gln Leu Gln Ile Lys Glu Ser Leu Glu Asp Leu Gln Glu Gln Leu 355 36aa gaa gct agt gat gaa aat caa aaa aga gaa ata gaa aag caa att s Glu Ala Ser Asp Glu Asn Gln Lys Arg Glu Ile Glu Lys Gln Ile 378tc aaa aaa aat gat gaa gaa ctt ttt aaa aat aaa gat cat aaa u Ile Lys Lys Asn Asp Glu Glu Leu Phe Lys Asn Lys Asp His Lys 385 39tta gat ctt aag caa gaa tta aat tct aaa gct tct agt aaa gaa a Leu Asp Leu Lys Gln Glu Leu Asn Ser Lys Ala Ser Ser Lys Glu 44att gaa ggc gaa gaa gag gat aaa gaa tta gat agt aaa aaa aat s Ile Glu Gly Glu Glu Glu Asp Lys Glu Leu Asp Ser Lys Lys Asn 423ag cct gtt tct gag gct gat aaa gta gat aaa att tcc aag tct u Glu Pro Val Ser Glu Ala Asp Lys Val Asp Lys Ile Ser Lys Ser 435 44ac aac aat gag gtt agt aaa tta tcc ccg tta gat gag cct tct tat n Asn Asn Glu Val Ser Lys Leu Ser Pro Leu Asp Glu Pro Ser Tyr 456ac att gat tcg aaa gag ggt gta gat aac aaa gat gtt gat ttg r Asp Ile Asp Ser Lys Glu Gly Val Asp Asn Lys Asp Val Asp Leu 465 478aa act aaa ccc caa gtt gaa agt caa cct act tcg tta aat gaa n Lys Thr Lys Pro Gln Val Glu Ser Gln Pro Thr Ser Leu Asn Glu 485 49ac ttg att gat gtg tct ata gat tcc agt aat cct gtc ttt tta gag p Leu Ile Asp Val Ser Ile Asp Ser Ser Asn Pro Val Phe Leu Glu 55atc gat ccg att aca aat tta gga acg ctt caa ctt att gat ttg l Ile Asp Pro Ile Thr Asn Leu Gly Thr Leu Gln Leu Ile Asp Leu 5525 aat acc ggt gtt aga ctt aaa gaa agt gct caa caa ggt att cag cga n Thr Gly Val Arg Leu Lys Glu Ser Ala Gln Gln Gly Ile Gln Arg 534ga att tat gaa cgt gaa aaa gat ttg gtt gtt att aaa ata gat r Gly Ile Tyr Glu Arg Glu Lys Asp Leu Val Val Ile Lys Ile Asp 545 556ga aaa gct aag ctt cag ata ctt gat aaa ctc gag aat tta aaa r Gly Lys Ala Lys Leu Gln Ile Leu Asp Lys Leu Glu Asn Leu Lys 565 57tg ata tca gag tct aat ttt gag att aat aaa aat tca tct ctt tat l Ile Ser Glu Ser Asn Phe Glu Ile Asn Lys Asn Ser Ser Leu Tyr 589ac tct aga atg att tta gta gtt gtt aag gac gat agt aat gct l Asp Ser Arg Met Ile Leu Val Val Val Lys Asp Asp Ser Asn Ala 595 6tgg aga ttg gct aaa ttt tct cct aaa aat tta gat gaa ttt att ctg p Arg Leu Ala Lys Phe Ser Pro Lys Asn Leu Asp Glu Phe Ile Leu 662aa aat aaa att ttg cct ttt act agc ttt gct gtg aga aag aat r Glu Asn Lys Ile Leu Pro Phe Thr Ser Phe Ala Val Arg Lys Asn 625 634tt tat ttg caa gat gaa ctt aaa agc tta gtt act tta gat gta e Ile Tyr Leu Gln Asp Glu Leu Lys Ser Leu Val Thr Leu Asp Val 645 65at act tta aaa aaa gtt aag ta n Thr Leu Lys Lys Val Lys 663 PRT Borrelia burgdorferi 78 Met Lys Lys Met Leu Leu Ile Phe Ser Phe Phe Leu Val Phe Leu Asn Phe Pro Leu Asn Ala Arg Glu Val Asp Lys Glu Lys Leu Lys Asp 2 Phe Val Asn Met Asp Leu Glu Phe Val Asn Tyr Lys Gly Pro Tyr Asp 35 4r Thr Asn Thr Tyr Glu Gln Ile Val Gly Ile Gly Glu Phe Leu Ala 5 Arg Pro Leu Ile Asn Ser Asn Ser Asn Ser Ser Tyr Tyr Gly Lys Tyr 65 7 Phe Val Asn Arg Phe Ile Asp Asp Gln Asp Lys Lys Ala Ser Val Asp 85 9e Phe Ser Ile Gly Ser Lys Ser Glu Leu Asp Ser Ile Leu Asn Leu Arg Ile Leu Thr Gly Tyr Leu Met Lys Ser Phe Asp Tyr Glu Arg Ser Ala Glu Leu Ile Ala Lys Ala Ile Thr Ile Tyr Asn Ala Val Arg Gly Asp Leu Asp Tyr Tyr Lys Glu Phe Tyr Ile Glu Ala Ser Leu Lys Ser Leu Thr Lys Glu Asn Ala Gly Leu Ser Arg Val Tyr Ser Trp Ala Gly Lys Thr Gln Ile Phe Ile Pro Leu Lys Lys Asn Ile Ser Gly Asn Val Glu Ser Asp Ile Asp Ile Asp Ser Leu Val Thr 2Lys Val Val Ala Ala Leu Leu Ser Glu Asn Glu Ser Gly Val Asn 222la Arg Asp Ile Thr Asp Ile Gln Gly Glu Thr His Lys Ala Asp 225 234sp Lys Ile Asp Ile Glu Leu Asp Asn Phe His Glu Ser Asp Ser 245 25sn Ile Thr Glu Thr Ile Glu Asn Leu Arg Asp Gln Leu Glu Lys Ala 267sp Glu Glu His Lys Lys Glu Ile Glu Ser Gln Val Asp Ala Lys 275 28ys Lys Gln Lys Glu Glu Leu Asp Lys Lys Ala Ile Asp Leu Asp Lys 29Gln Gln Lys Leu Asp Phe Ala Glu Asp Asn Leu Asp Ile Gln Arg 33Asp Thr Val Arg Glu Lys Leu Gln Glu Asn Ile Asn Glu Thr Asn Lys 325 33lu Lys Asn Leu Pro Lys Pro Gly Asp Val Ser Ser Pro Lys Val Asp 345ln Leu Gln Ile Lys Glu Ser Leu Glu Asp Leu Gln Glu Gln Leu 355 36ys Glu Ala Ser Asp Glu Asn Gln Lys Arg Glu Ile Glu Lys Gln Ile 378le Lys Lys Asn Asp Glu Glu Leu Phe Lys Asn Lys Asp His Lys 385 39Leu Asp Leu Lys Gln Glu Leu Asn Ser Lys Ala Ser Ser Lys Glu 44Ile Glu Gly Glu Glu Glu Asp Lys Glu Leu Asp Ser Lys Lys Asn 423lu Pro Val Ser Glu Ala Asp Lys Val Asp Lys Ile Ser Lys Ser 435 44sn Asn Asn Glu Val Ser Lys Leu Ser Pro Leu Asp Glu Pro Ser Tyr 456sp Ile Asp Ser Lys Glu Gly Val Asp Asn Lys Asp Val Asp Leu 465 478ys Thr Lys Pro Gln Val Glu Ser Gln Pro Thr Ser Leu Asn Glu 485 49sp Leu Ile Asp Val Ser Ile Asp Ser Ser Asn Pro Val Phe Leu Glu 55Ile Asp Pro Ile Thr Asn Leu Gly Thr Leu Gln Leu Ile Asp Leu 5525 Asn Thr Gly Val Arg Leu Lys Glu Ser Ala Gln Gln Gly Ile Gln Arg 534ly Ile Tyr Glu Arg Glu Lys Asp Leu Val Val Ile Lys Ile Asp 545 556ly Lys Ala Lys Leu Gln Ile Leu Asp Lys Leu Glu Asn Leu Lys 565 57al Ile Ser Glu Ser Asn Phe Glu Ile Asn Lys Asn Ser Ser Leu Tyr 589sp Ser Arg Met Ile Leu Val Val Val Lys Asp Asp Ser Asn Ala 595 6Trp Arg Leu Ala Lys Phe Ser Pro Lys Asn Leu Asp Glu Phe Ile Leu 662lu Asn Lys Ile Leu Pro Phe Thr Ser Phe Ala Val Arg Lys Asn 625 634le Tyr Leu Gln Asp Glu Leu Lys Ser Leu Val Thr Leu Asp Val 645 65sn Thr Leu Lys Lys Val Lys 66 DNA Artificial Sequence Oligonucleotide Primer 79 ccggtcaccc catggctgct ttaaagtctt ta 32 8A Artificial Sequence Oligonucleotide Primer 8caccc catgaatctt gataaagctc ag 32 8A Artificial Sequence Oligonucleotide Primer 8caccc catggatgaa aagcttttaa aaagt 35 82 33 DNA Artificial Sequence Oligonucleotide Primer 82 ccggtcaccc ccatggttga gaaattagat aag 33 83 32 DNA Artificial Sequence Oligonucleotide Primer 83 ttggatccgg tgacccttaa

ctttttttaa ag 32 84 38 DNA Artificial Sequence Oligonucleotide Primer 84 aaagtagaag tttttgaatc ccattttcca gttttttt 38 85 825 DNA Artificial Sequence CDS (25) Chimeric nucleic acid 85 atg aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gat gaa aaa aat agc gtt tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 gat tta cct ggt gga atg aca gtt ctt gta agt aaa gaa aaa gac aaa Leu Pro Gly Gly Met Thr Val Leu Val Ser Lys Glu Lys Asp Lys 35 4c ggt aaa tac agt cta gag gca aca gta gac aag ctt gag ctt aaa Gly Lys Tyr Ser Leu Glu Ala Thr Val Asp Lys Leu Glu Leu Lys 5 gga act tct gat aaa aac aac ggt tct gga aca ctt gaa ggt gaa aaa 24hr Ser Asp Lys Asn Asn Gly Ser Gly Thr Leu Glu Gly Glu Lys 65 7 act gac aaa agt aaa gta aaa tta aca att gct gat gac cta agt caa 288 Thr Asp Lys Ser Lys Val Lys Leu Thr Ile Ala Asp Asp Leu Ser Gln 85 9t aaa ttt gaa att ttc aaa gaa gat gcc aaa aca tta gta tca aaa 336 Thr Lys Phe Glu Ile Phe Lys Glu Asp Ala Lys Thr Leu Val Ser Lys gta acc ctt aaa gac aag tca tca aca gaa gaa aaa ttc aac gaa 384 Lys Val Thr Leu Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu ggt gaa aca tct gaa aaa aca ata gta aga gca aat gga acc aga 432 Lys Gly Glu Thr Ser Glu Lys Thr Ile Val Arg Ala Asn Gly Thr Arg gaa tac aca gac ata aaa agc gat gga tcc gga aaa gct aaa gaa 48lu Tyr Thr Asp Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu gtt tta aaa gac ttt act ctt gaa gga act cta gct gct gac ggc aaa 528 Val Leu Lys Asp Phe Thr Leu Glu Gly Thr Leu Ala Ala Asp Gly Lys aca ttg aaa gtt aca gaa ggc act gtt gtt tta agc aag aac att 576 Thr Thr Leu Lys Val Thr Glu Gly Thr Val Val Leu Ser Lys Asn Ile aaa tcc gga gaa ata aca gtt gca ctt gat gac tct gac act act 624 Leu Lys Ser Gly Glu Ile Thr Val Ala Leu Asp Asp Ser Asp Thr Thr 2gct act aaa aaa act gga aaa tgg gat tca aaa act tct act tta 672 Gln Ala Thr Lys Lys Thr Gly Lys Trp Asp Ser Lys Thr Ser Thr Leu 222tt agt gtt aac agc aaa aaa act aca caa ctt gtg ttt act aaa 72le Ser Val Asn Ser Lys Lys Thr Thr Gln Leu Val Phe Thr Lys 225 234ac aca ata act gta aaa caa tac gac tcc gca ggt acc aat tta 768 Gln Tyr Thr Ile Thr Val Lys Gln Tyr Asp Ser Ala Gly Thr Asn Leu 245 25aa ggc aca gca gtc gaa att aaa aca ctt gat gaa ctt aaa aac gct 8Gly Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala 267aa taa 825 Leu Lys * 86 274 PRT Artificial Sequence Chimeric Protein 86 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 Asp Leu Pro Gly Gly Met Thr Val Leu Val Ser Lys Glu Lys Asp Lys 35 4p Gly Lys Tyr Ser Leu Glu Ala Thr Val Asp Lys Leu Glu Leu Lys 5 Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Thr Leu Glu Gly Glu Lys 65 7 Thr Asp Lys Ser Lys Val Lys Leu Thr Ile Ala Asp Asp Leu Ser Gln 85 9r Lys Phe Glu Ile Phe Lys Glu Asp Ala Lys Thr Leu Val Ser Lys Val Thr Leu Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Gly Glu Thr Ser Glu Lys Thr Ile Val Arg Ala Asn Gly Thr Arg Glu Tyr Thr Asp Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys Asp Phe Thr Leu Glu Gly Thr Leu Ala Ala Asp Gly Lys Thr Leu Lys Val Thr Glu Gly Thr Val Val Leu Ser Lys Asn Ile Lys Ser Gly Glu Ile Thr Val Ala Leu Asp Asp Ser Asp Thr Thr 2Ala Thr Lys Lys Thr Gly Lys Trp Asp Ser Lys Thr Ser Thr Leu 222le Ser Val Asn Ser Lys Lys Thr Thr Gln Leu Val Phe Thr Lys 225 234yr Thr Ile Thr Val Lys Gln Tyr Asp Ser Ala Gly Thr Asn Leu 245 25lu Gly Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala 267ys 87 35 DNA Artificial Sequence Oligonucleotide Primer 87 aaagtagaag tttttgaatt ccaagctgca gtttt 35 88 822 DNA Artificial Sequence CDS (22) Chimeric nucleic acid 88 atg aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gat gaa aaa aac agc gct tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Ala Ser Val 2 gat ttg cct ggt gag atg aaa gtt ctt gta agt aaa gaa aaa gac aaa Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asp Lys 35 4c ggt aag tac agt cta aag gca aca gta gac aag att gag cta aaa Gly Lys Tyr Ser Leu Lys Ala Thr Val Asp Lys Ile Glu Leu Lys 5 gga act tct gat aaa gac aat ggt tct gga gtg ctt gaa ggt aca aaa 24hr Ser Asp Lys Asp Asn Gly Ser Gly Val Leu Glu Gly Thr Lys 65 7 gat gac aaa agt aaa gca aaa tta aca att gct gac gat cta agt aaa 288 Asp Asp Lys Ser Lys Ala Lys Leu Thr Ile Ala Asp Asp Leu Ser Lys 85 9c aca ttc gaa ctt tta aaa gaa gat ggc aaa aca tta gtg tca aga 336 Thr Thr Phe Glu Leu Leu Lys Glu Asp Gly Lys Thr Leu Val Ser Arg gta agt tct aga gac aaa aca tca aca gat gaa atg ttc aat gaa 384 Lys Val Ser Ser Arg Asp Lys Thr Ser Thr Asp Glu Met Phe Asn Glu ggt gaa ttg tct gca aaa acc atg aca aga gaa aat gga acc aaa 432 Lys Gly Glu Leu Ser Ala Lys Thr Met Thr Arg Glu Asn Gly Thr Lys gaa tat aca gaa atg aaa agc gat gga acc gga aaa gct aaa gaa 48lu Tyr Thr Glu Met Lys Ser Asp Gly Thr Gly Lys Ala Lys Glu gtt tta aaa aag ttt act ctt gaa gga aaa gta gct aat gat aaa gta 528 Val Leu Lys Lys Phe Thr Leu Glu Gly Lys Val Ala Asn Asp Lys Val ttg gaa gta aaa gaa gga acc gtt act tta agt aag gaa att gca 576 Thr Leu Glu Val Lys Glu Gly Thr Val Thr Leu Ser Lys Glu Ile Ala tct gga gaa gta aca gtt gct ctt aat gac act aac act act cag 624 Lys Ser Gly Glu Val Thr Val Ala Leu Asn Asp Thr Asn Thr Thr Gln 2act aaa aaa act ggc gca tgg gat tca aaa act tct act tta aca 672 Ala Thr Lys Lys Thr Gly Ala Trp Asp Ser Lys Thr Ser Thr Leu Thr 222gt gtt aac agc aaa aaa act aca caa ctt gtg ttt act aaa caa 72er Val Asn Ser Lys Lys Thr Thr Gln Leu Val Phe Thr Lys Gln 225 234ca ata act gta aaa caa tac gac tcc gca ggt acc aat tta gaa 768 Tyr Thr Ile Thr Val Lys Gln Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25gc aca gca gtc gaa att aaa aca ctt gat gaa ctt aaa aac gct tta 8Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu 267aa 822 Lys * 89 273 PRT Artificial Sequence Chimeric Protein 89 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Ala Ser Val 2 Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asp Lys 35 4p Gly Lys Tyr Ser Leu Lys Ala Thr Val Asp Lys Ile Glu Leu Lys 5 Gly Thr Ser Asp Lys Asp Asn Gly Ser Gly Val Leu Glu Gly Thr Lys 65 7 Asp Asp Lys Ser Lys Ala Lys Leu Thr Ile Ala Asp Asp Leu Ser Lys 85 9r Thr Phe Glu Leu Leu Lys Glu Asp Gly Lys Thr Leu Val Ser Arg Val Ser Ser Arg Asp Lys Thr Ser Thr Asp Glu Met Phe Asn Glu Gly Glu Leu Ser Ala Lys Thr Met Thr Arg Glu Asn Gly Thr Lys Glu Tyr Thr Glu Met Lys Ser Asp Gly Thr Gly Lys Ala Lys Glu Val Leu Lys Lys Phe Thr Leu Glu Gly Lys Val Ala Asn Asp Lys Val Leu Glu Val Lys Glu Gly Thr Val Thr Leu Ser Lys Glu Ile Ala Ser Gly Glu Val Thr Val Ala Leu Asn Asp Thr Asn Thr Thr Gln 2Thr Lys Lys Thr Gly Ala Trp Asp Ser Lys Thr Ser Thr Leu Thr 222er Val Asn Ser Lys Lys Thr Thr Gln Leu Val Phe Thr Lys Gln 225 234hr Ile Thr Val Lys Gln Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25ly Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu 267A Artificial Sequence Oligonucleotide Primer 9ggaag tttttgaatt ccaagctgca gttttttt 38 9NA Artificial Sequence CDS (22) Chimeric nucleic acid 9aa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gac gag aaa aac agc gtt tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 gat ttg cct ggt gaa atg aaa gtt ctt gta agc aaa gaa aaa aac aaa Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys 35 4c ggc aag tac gat cta att gca aca gta gac aag ctt gag ctt aaa Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys 5 gga act tct gat aaa aac aat gga tct gga gta ctt gaa ggc gta aaa 24hr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 gct gac aaa agt aaa gta aaa tta aca att tct gac gat cta ggt caa 288 Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln 85 9c aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca cta gta tca aaa 336 Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys gta act tcc aaa gac aag tca tca aca gaa gaa aaa ttc aat gaa 384 Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu ggt gaa gta tct gaa aaa ata ata aca aga gca gac gga acc aga 432 Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg gaa tac aca gga att aaa agc gat gga tct gga aaa gct aaa gag 48lu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu gtt tta aaa ggc tat gtt ctt gaa gga act cta act gct gaa aaa aca 528 Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr ttg gtg gtt aaa gaa gga act gtt act tta agc aaa aat att tca 576 Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser tct ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt gct 624 Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2act aaa aaa act gca gct tgg aat tca aaa act tcc act tta aca 672 Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr 222gt gtg aat agc caa aaa acc aaa aac ctt gta ttc aca aaa gaa 72er Val Asn Ser Gln Lys Thr Lys Asn Leu Val Phe Thr Lys Glu 225 234ca ata aca gta caa aaa tac gac tca gca ggc acc aat cta gaa 768 Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25gc aaa gca gtc gaa att aca aca ctt aaa gaa ctt aaa aac gct tta 8Lys Ala Val Glu Ile Thr Thr Leu Lys Glu Leu Lys Asn Ala Leu 267aa 822 Lys * 92 273 PRT Artificial Sequence Chimeric Protein 92 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys 35 4p Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys 5 Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln 85 9r Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr 222er Val Asn Ser Gln Lys Thr Lys Asn Leu Val Phe Thr Lys Glu 225 234hr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25ly Lys Ala Val Glu Ile Thr Thr Leu Lys Glu Leu Lys Asn Ala Leu 2673 35 DNA Artificial Sequence Oligonucleotide Primer 93 taaagttgaa gtgcctgcat tccaagctgc agttt 35 94 8Artificial Sequence CDS (meric nucleic acid 94 atg aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gac gag aaa aac agc gtt tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 gat ttg cct ggt gaa atg aaa gtt ctt gta agc aaa gaa aaa aac aaa Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys 35 4c ggc aag tac gat cta att gca aca gta gac aag ctt gag ctt aaa Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys 5 gga act tct gat aaa aac aat gga tct gga gta ctt gaa ggc gta aaa 24hr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 gct gac aaa agt aaa gta aaa tta aca att tct gac gat cta ggt caa 288 Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu

Gly Gln 85 9c aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca cta gta tca aaa 336 Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys gta act tcc aaa gac aag tca tca aca gaa gaa aaa ttc aat gaa 384 Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu ggt gaa gta tct gaa aaa ata ata aca aga gca gac gga acc aga 432 Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg gaa tac aca gga att aaa agc gat gga tct gga aaa gct aaa gag 48lu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu gtt tta aaa ggc tat gtt ctt gaa gga act cta act gct gaa aaa aca 528 Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr ttg gtg gtt aaa gaa gga act gtt act tta agc aaa aat att tca 576 Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser tct ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt gct 624 Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2act aaa aaa act gca gct tgg aat gca ggc act tca act tta aca 672 Ala Thr Lys Lys Thr Ala Ala Trp Asn Ala Gly Thr Ser Thr Leu Thr 222ct gta aac aac aaa aaa act aaa gcc ctt gta ttt aca aaa caa 72hr Val Asn Asn Lys Lys Thr Lys Ala Leu Val Phe Thr Lys Gln 225 234ca att aca tca caa aaa tac gac tca gca gga acc aac ttg gaa 768 Asp Thr Ile Thr Ser Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25gc aca gca gtc gaa att aaa aca ctt gat gaa ctt aaa aac gct tta 8Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu 26795 273 PRT Artificial Sequence Chimeric Protein 95 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys 35 4p Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys 5 Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln 85 9r Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2Thr Lys Lys Thr Ala Ala Trp Asn Ala Gly Thr Ser Thr Leu Thr 222hr Val Asn Asn Lys Lys Thr Lys Ala Leu Val Phe Thr Lys Gln 225 234hr Ile Thr Ser Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25ly Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu 2676 3rtificial Sequence Oligonucleotide Primer 96 ccccagattt tgaaatcttg cttaaaacaa c 32 DNA Artificial Sequence CDS (22) Chimeric nucleic acid 97 atg aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gat gaa aaa aat agc gtt tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 gat tta cct ggt gga atg aca gtt ctt gta agt aaa gaa aaa gac aaa Leu Pro Gly Gly Met Thr Val Leu Val Ser Lys Glu Lys Asp Lys 35 4c ggt aaa tac agt cta gag gca aca gta gac aag ctt gag ctt aaa Gly Lys Tyr Ser Leu Glu Ala Thr Val Asp Lys Leu Glu Leu Lys 5 gga act tct gat aaa aac aac ggt tct gga aca ctt gaa ggt gaa aaa 24hr Ser Asp Lys Asn Asn Gly Ser Gly Thr Leu Glu Gly Glu Lys 65 7 act gac aaa agt aaa gta aaa tta aca att gct gat gac cta agt caa 288 Thr Asp Lys Ser Lys Val Lys Leu Thr Ile Ala Asp Asp Leu Ser Gln 85 9t aaa ttt gaa att ttc aaa gaa gat gcc aaa aca tta gta tca aaa 336 Thr Lys Phe Glu Ile Phe Lys Glu Asp Ala Lys Thr Leu Val Ser Lys gta acc ctt aaa gac aag tca tca aca gaa gaa aaa ttc aac gaa 384 Lys Val Thr Leu Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu ggt gaa aca tct gaa aaa aca ata gta aga gca aat gga acc aga 432 Lys Gly Glu Thr Ser Glu Lys Thr Ile Val Arg Ala Asn Gly Thr Arg gaa tac aca gac ata aaa agc gat gga tcc gga aaa gct aaa gaa 48lu Tyr Thr Asp Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu gtt tta aaa gac ttt act ctt gaa gga act cta gct gct gac ggc aaa 528 Val Leu Lys Asp Phe Thr Leu Glu Gly Thr Leu Ala Ala Asp Gly Lys aca ttg aaa gtt aca gaa ggc act gtt gtt tta agc aag att tca 576 Thr Thr Leu Lys Val Thr Glu Gly Thr Val Val Leu Ser Lys Ile Ser tct ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt gct 624 Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2act aaa aaa act gca gct tgg aat tca aaa act tcc act tta aca 672 Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr 222gt gtg aat agc caa aaa acc aaa aac ctt gta ttc aca aaa gaa 72er Val Asn Ser Gln Lys Thr Lys Asn Leu Val Phe Thr Lys Glu 225 234ca ata aca gta caa aaa tac gac tca gca ggc acc aat cta gaa 768 Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25gc aaa gca gtc gaa att aca aca ctt aaa gaa ctt aaa aac gct tta 8Lys Ala Val Glu Ile Thr Thr Leu Lys Glu Leu Lys Asn Ala Leu 267aa 822 Lys * 98 273 PRT Artificial Sequence Chimeric Protein 98 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 Asp Leu Pro Gly Gly Met Thr Val Leu Val Ser Lys Glu Lys Asp Lys 35 4p Gly Lys Tyr Ser Leu Glu Ala Thr Val Asp Lys Leu Glu Leu Lys 5 Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Thr Leu Glu Gly Glu Lys 65 7 Thr Asp Lys Ser Lys Val Lys Leu Thr Ile Ala Asp Asp Leu Ser Gln 85 9r Lys Phe Glu Ile Phe Lys Glu Asp Ala Lys Thr Leu Val Ser Lys Val Thr Leu Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Gly Glu Thr Ser Glu Lys Thr Ile Val Arg Ala Asn Gly Thr Arg Glu Tyr Thr Asp Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys Asp Phe Thr Leu Glu Gly Thr Leu Ala Ala Asp Gly Lys Thr Leu Lys Val Thr Glu Gly Thr Val Val Leu Ser Lys Ile Ser Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr 222er Val Asn Ser Gln Lys Thr Lys Asn Leu Val Phe Thr Lys Glu 225 234hr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25ly Lys Ala Val Glu Ile Thr Thr Leu Lys Glu Leu Lys Asn Ala Leu 2679 32 DNA Artificial Sequence Oligonucleotide Primer 99 caagtctggt tccaatttgc tcttgttatt at 32 DNA Artificial Sequence CDS (22) Chimeric nucleic acid aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gat gaa aaa aat agc gtt tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 gat tta cct ggt gga atg aca gtt ctt gta agt aaa gaa aaa gac aaa Leu Pro Gly Gly Met Thr Val Leu Val Ser Lys Glu Lys Asp Lys 35 4c ggt aaa tac agt cta gag gca aca gta gac aag ctt gag ctt aaa Gly Lys Tyr Ser Leu Glu Ala Thr Val Asp Lys Leu Glu Leu Lys 5 gga act tct gat aaa aac aac ggt tct gga aca ctt gaa ggt gaa aaa 24hr Ser Asp Lys Asn Asn Gly Ser Gly Thr Leu Glu Gly Glu Lys 65 7 act gac aaa agt aaa gta aaa tta aca att gct gat gac cta agt caa 288 Thr Asp Lys Ser Lys Val Lys Leu Thr Ile Ala Asp Asp Leu Ser Gln 85 9t aaa ttt gaa att ttc aaa gaa gat gcc aaa aca tta gta tca aaa 336 Thr Lys Phe Glu Ile Phe Lys Glu Asp Ala Lys Thr Leu Val Ser Lys gta acc ctt aaa gac aag tca tca aca gaa gaa aaa ttc aac gaa 384 Lys Val Thr Leu Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu ggt gaa aca tct gaa aaa aca ata gta aga gca aat gga acc aga 432 Lys Gly Glu Thr Ser Glu Lys Thr Ile Val Arg Ala Asn Gly Thr Arg gaa tac aca gac ata aaa agc gat gga tcc gga aaa gct aaa gaa 48lu Tyr Thr Asp Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu gtt tta aaa gac ttt act ctt gaa gga act cta gct gct gac ggc aaa 528 Val Leu Lys Asp Phe Thr Leu Glu Gly Thr Leu Ala Ala Asp Gly Lys aca ttg aaa gtt aca gaa ggc act gtt gtt tta agc aag att tca 576 Thr Thr Leu Lys Val Thr Glu Gly Thr Val Val Leu Ser Lys Ile Ser tct ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt gct 624 Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2act aaa aaa act gca gct tgg aat tca aaa act tcc act tta aca 672 Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr 222gt gtg aat agc caa aaa acc aaa aac ctt gta ttc aca aaa gaa 72er Val Asn Ser Gln Lys Thr Lys Asn Leu Val Phe Thr Lys Glu 225 234ca ata aca gta caa aaa tac gac tca gca ggc acc aat cta gaa 768 Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25gc aaa gca gtc gaa att aca aca ctt aaa gaa ctt aaa aac gct tta 8Lys Ala Val Glu Ile Thr Thr Leu Lys Glu Leu Lys Asn Ala Leu 267aa 822 Lys * PRT Artificial Sequence Chimeric Protein Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 Asp Leu Pro Gly Gly Met Thr Val Leu Val Ser Lys Glu Lys Asp Lys 35 4p Gly Lys Tyr Ser Leu Glu Ala Thr Val Asp Lys Leu Glu Leu Lys 5 Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Thr Leu Glu Gly Glu Lys 65 7 Thr Asp Lys Ser Lys Val Lys Leu Thr Ile Ala Asp Asp Leu Ser Gln 85 9r Lys Phe Glu Ile Phe Lys Glu Asp Ala Lys Thr Leu Val Ser Lys Val Thr Leu Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Gly Glu Thr Ser Glu Lys Thr Ile Val Arg Ala Asn Gly Thr Arg Glu Tyr Thr Asp Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys Asp Phe Thr Leu Glu Gly Thr Leu Ala Ala Asp Gly Lys Thr Leu Lys Val Thr Glu Gly Thr Val Val Leu Ser Lys Ile Ser Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr 222er Val Asn Ser Gln Lys Thr Lys Asn Leu Val Phe Thr Lys Glu 225 234hr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25ly Lys Ala Val Glu Ile Thr Thr Leu Lys Glu Leu Lys Asn Ala Leu 267NA Artificial Sequence Oligonucleotide Primer aaagtgc tagtactgtc attccaagct gcagtttttt t 422 DNA Artificial Sequence CDS (22) Chimeric nucleic acid aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gac gag aaa aac agc gtt tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 gat ttg cct ggt gaa atg aaa gtt ctt gta agc aaa gaa aaa aac aaa Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys 35 4c ggc aag tac gat cta att gca aca gta gac aag ctt gag ctt aaa Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys 5 gga act tct gat aaa aac aat gga tct gga gta ctt gaa ggc gta aaa 24hr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 gct gac aaa agt aaa gta aaa tta aca att tct gac gat cta ggt caa 288 Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln 85 9c aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca cta gta tca aaa 336 Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys gta act tcc aaa gac aag tca tca aca gaa gaa aaa ttc aat gaa 384 Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu ggt gaa gta tct gaa aaa ata ata aca aga gca gac gga acc aga 432 Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg gaa tac aca gga att aaa agc gat gga tct gga aaa gct aaa gag 48lu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu gtt tta aaa ggc tat gtt ctt gaa gga act cta act gct gaa aaa aca 528 Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr ttg gtg gtt aaa gaa gga act gtt act tta agc aaa aat att tca 576 Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser tct ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt gct 624 Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser

Ser Ala 2act aaa aaa act gca gct tgg aat gac agt act agc act tta aca 672 Ala Thr Lys Lys Thr Ala Ala Trp Asn Asp Ser Thr Ser Thr Leu Thr 222gt gct gac agc aaa aaa act aaa gat ttg gtg ttc tta aca gat 72er Ala Asp Ser Lys Lys Thr Lys Asp Leu Val Phe Leu Thr Asp 225 234ca att aca gta caa caa tac aac aca gct gga acc agc cta gaa 768 Gly Thr Ile Thr Val Gln Gln Tyr Asn Thr Ala Gly Thr Ser Leu Glu 245 25ga tca gca agt gaa att aaa aat ctt tca gag ctt aaa aac gct tta 8Ser Ala Ser Glu Ile Lys Asn Leu Ser Glu Leu Lys Asn Ala Leu 267aa 822 Lys * PRT Artificial Sequence Chimeric Protein Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys 35 4p Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys 5 Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln 85 9r Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2Thr Lys Lys Thr Ala Ala Trp Asn Asp Ser Thr Ser Thr Leu Thr 222er Ala Asp Ser Lys Lys Thr Lys Asp Leu Val Phe Leu Thr Asp 225 234hr Ile Thr Val Gln Gln Tyr Asn Thr Ala Gly Thr Ser Leu Glu 245 25ly Ser Ala Ser Glu Ile Lys Asn Leu Ser Glu Leu Lys Asn Ala Leu 267NA Artificial Sequence Oligonucleotide Primer agatgta atcccatccg ccatttttaa agcgttttt 39 rtificial Sequence CDS (4meric nucleic acid aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gac gag aaa aac agc gtt tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 gat ttg cct ggt gaa atg aaa gtt ctt gta agc aaa gaa aaa aac aaa Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys 35 4c ggc aag tac gat cta att gca aca gta gac aag ctt gag ctt aaa Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys 5 gga act tct gat aaa aac aat gga tct gga gta ctt gaa ggc gta aaa 24hr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 gct gac aaa agt aaa gta aaa tta aca att tct gac gat cta ggt caa 288 Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln 85 9c aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca cta gta tca aaa 336 Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys gta act tcc aaa gac aag tca tca aca gaa gaa aaa ttc aat gaa 384 Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu ggt gaa gta tct gaa aaa ata ata aca aga gca gac gga acc aga 432 Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg gaa tac aca gga att aaa agc gat gga tct gga aaa gct aaa gag 48lu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu gtt tta aaa ggc tat gtt ctt gaa gga act cta act gct gaa aaa aca 528 Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr ttg gtg gtt aaa gaa gga act gtt act tta agc aaa aat att tca 576 Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser tct ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt gct 624 Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2act aaa aaa act gca gct tgg aat gac agt act agc act tta aca 672 Ala Thr Lys Lys Thr Ala Ala Trp Asn Asp Ser Thr Ser Thr Leu Thr 222gt gct gac agc aaa aaa act aaa gat ttg gtg ttc tta aca gat 72er Ala Asp Ser Lys Lys Thr Lys Asp Leu Val Phe Leu Thr Asp 225 234ca att aca gta caa caa tac aac aca gct gga acc agc cta gaa 768 Gly Thr Ile Thr Val Gln Gln Tyr Asn Thr Ala Gly Thr Ser Leu Glu 245 25ga tca gca agt gaa att aaa aat ctt tca gag ctt aaa aac gct tta 8Ser Ala Ser Glu Ile Lys Asn Leu Ser Glu Leu Lys Asn Ala Leu 267tg gct aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 864 Lys Met Ala Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser 275 28ct gat gag tct gtt aaa ggg cct aat ctt aca gaa ata agt aaa aaa 9Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 29acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg 96hr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 33ttg ctg tca tct ata gat gaa att gct gct aaa gct att ggt aaa aaa u Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 325 33ta cac caa aat aat ggt ttg gat acc gaa tat aat cac aat gga tca e His Gln Asn Asn Gly Leu Asp Thr Glu Tyr Asn His Asn Gly Ser 345ta gcg gga cgt tat gca ata tca acc cta ata aaa caa aaa tta u Leu Ala Gly Arg Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 355 36at gga ttg aaa aat gaa gga tta aag gaa aaa att gat gcg gct aag p Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys 378gt tct gaa aca ttt act aat aaa tta aaa gca aaa cac aca gat s Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Ala Lys His Thr Asp 385 39ggt aaa gaa ggt gtt act gat gct gat gca aaa gaa gcc att tta u Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu 44aca aat ggt act aaa act aaa ggt gct gaa gaa ctt gga aaa tta s Thr Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu 423aa tca gta gag gtc ttg tca aaa gca gct aaa gag atg ctt gct e Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala 435 44at tca gtt aaa gag ctt aca agc cct gtt gtg gca gaa agt cca aaa n Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys 456ct taa s Pro * 465 PRT Artificial Sequence Chimeric Protien Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys 35 4p Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys 5 Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln 85 9r Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2Thr Lys Lys Thr Ala Ala Trp Asn Asp Ser Thr Ser Thr Leu Thr 222er Ala Asp Ser Lys Lys Thr Lys Asp Leu Val Phe Leu Thr Asp 225 234hr Ile Thr Val Gln Gln Tyr Asn Thr Ala Gly Thr Ser Leu Glu 245 25ly Ser Ala Ser Glu Ile Lys Asn Leu Ser Glu Leu Lys Asn Ala Leu 267et Ala Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser 275 28la Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 29Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 33Leu Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 325 33le His Gln Asn Asn Gly Leu Asp Thr Glu Tyr Asn His Asn Gly Ser 345eu Ala Gly Arg Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 355 36sp Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys 378ys Ser Glu Thr Phe Thr Asn Lys Leu Lys Ala Lys His Thr Asp 385 39Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu 44Thr Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu 423lu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala 435 44sn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys 456ro 465 DNA Artificial Sequence Oligonucleotide Primer gctaaca ttttgcttag gtttttttgg actttc 36 rtificial Sequence CDS (4meric Nucleic Acid aaa aag aat aca tta agt gcg ata tta atg act tta ttt tta ttt 48 Met Lys Lys Asn Thr Leu Ser Ala Ile Leu Met Thr Leu Phe Leu Phe tct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 96 Ile Ser Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser 2 gct gat gag tct gtt aaa ggg cct aat ctt aca gaa ata agt aaa aaa Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 35 4t acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 5 ttg ctg tca tct ata gat gaa att gct gct aaa gct att ggt aaa aaa 24eu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 65 7 ata cac caa aat aat ggt ttg gat acc gaa tat aat cac aat gga tca 288 Ile His Gln Asn Asn Gly Leu Asp Thr Glu Tyr Asn His Asn Gly Ser 85 9g tta gcg gga cgt tat gca ata tca acc cta ata aaa caa aaa tta 336 Leu Leu Ala Gly Arg Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu gga ttg aaa aat gaa gga tta aag gaa aaa att gat gcg gct aag 384 Asp Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys tgt tct gaa aca ttt act aat aaa tta aaa gca aaa cac aca gat 432 Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Ala Lys His Thr Asp ggt aaa gaa ggt gtt act gat gct gat gca aaa gaa gcc att tta 48ly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu aaa aca aat ggt act aaa act aaa ggt gct gaa gaa ctt gga aaa tta 528 Lys Thr Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu gaa tca gta gag gtc ttg tca aaa gca gct aaa gag atg ctt gct 576 Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala tca gtt aaa gag ctt aca agc cct gtt gtg gca gaa agt cca aaa 624 Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys 2cct aag caa aat gtt agc agc ctt gac gag aaa aac agc gtt tca 672 Lys Pro Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser 222at ttg cct ggt gaa atg aaa gtt ctt gta agc aaa gaa aaa aac 72sp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn 225 234ac ggc aag tac gat cta att gca aca gta gac aag ctt gag ctt 768 Lys Asp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu 245 25aa gga act tct gat aaa aac aat gga tct gga gta ctt gaa ggc gta 8Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val 267ct gac aaa agt aaa gta aaa tta aca att tct gac gat cta ggt 864 Lys Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly 275 28aa acc aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca cta gta tca 9Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser 29aaa gta act tcc aaa gac aag tca tca aca gaa gaa aaa ttc aat 96ys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn 33gaa aaa ggt gaa gta tct gaa aaa ata ata aca aga gca gac gga acc u Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr 325 33ga ctt gaa tac aca gga att aaa agc gat gga tct gga aaa gct aaa g Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys 345tt tta aaa ggc tat gtt ctt gaa gga act cta act gct gaa aaa u Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys 355 36ca aca ttg gtg gtt aaa gaa gga act gtt act tta agc aaa aat att r Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile 378aa tct ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt r Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser 385 39gct act aaa aaa act gca gct tgg aat gac agt act agc act tta a Ala Thr Lys Lys Thr Ala Ala Trp Asn Asp Ser Thr Ser Thr Leu 44att agt gct gac agc aaa aaa act aaa gat ttg gtg ttc tta aca r Ile Ser Ala Asp Ser Lys Lys Thr Lys Asp Leu Val Phe Leu Thr 423gt aca att aca gta caa caa tac aac aca gct gga acc agc cta p Gly Thr Ile Thr Val Gln Gln Tyr Asn Thr Ala Gly Thr Ser Leu 435 44aa gga tca gca agt gaa att aaa aat ctt tca gag ctt aaa aac gct u Gly Ser Ala Ser Glu Ile Lys Asn Leu Ser Glu Leu Lys Asn Ala 456aa taa u Lys * 465

PRT Artificial Sequence Chimeric Protein Lys Lys Asn Thr Leu Ser Ala Ile Leu Met Thr Leu Phe Leu Phe Ser Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser 2 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 35 4e Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 5 Leu Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 65 7 Ile His Gln Asn Asn Gly Leu Asp Thr Glu Tyr Asn His Asn Gly Ser 85 9u Leu Ala Gly Arg Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Ala Lys His Thr Asp Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu Lys Thr Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys 2Pro Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser 222sp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn 225 234sp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu 245 25ys Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val 267la Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly 275 28ln Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser 29Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn 33Glu Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr 325 33rg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys 345al Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys 355 36hr Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile 378ys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser 385 39Ala Thr Lys Lys Thr Ala Ala Trp Asn Asp Ser Thr Ser Thr Leu 44Ile Ser Ala Asp Ser Lys Lys Thr Lys Asp Leu Val Phe Leu Thr 423ly Thr Ile Thr Val Gln Gln Tyr Asn Thr Ala Gly Thr Ser Leu 435 44lu Gly Ser Ala Ser Glu Ile Lys Asn Leu Ser Glu Leu Lys Asn Ala 456ys 465 rtificial Sequence CDS (7meric Nucleic Acid caa aat gtt agc agc ctt gac gag aaa aac agc gtt tca gta gat 48 Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val Asp cct ggt gaa atg aaa gtt ctt gta agc aaa gaa aaa aac aaa gac 96 Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys Asp 2 ggc aag tac gat cta att gca aca gta gac aag ctt gag ctt aaa gga Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys Gly 35 4t tct gat aaa aac aat gga tct gga gta ctt gaa ggc gta aaa gct Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys Ala 5 gac aaa agt aaa gta aaa tta aca att tct gac gat cta ggt caa acc 24ys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln Thr 65 7 aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca cta gta tca aaa aaa 288 Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Lys 85 9a act tcc aaa gac aag tca tca aca gaa gaa aaa ttc aat gaa aaa 336 Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Lys gaa gta tct gaa aaa ata ata aca aga gca gac gga acc aga ctt 384 Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg Leu tac aca gga att aaa agc gat gga tct gga aaa gct aaa gag gtt 432 Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val aaa ggc tat gtt ctt gaa gga act cta act gct gaa aaa aca aca 48ys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr Thr ttg gtg gtt aaa gaa gga act gtt act tta agc aaa aat att tca aaa 528 Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser Lys ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt gct gct 576 Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala Ala aaa aaa act gca gct tgg aat tca ggc act tca act tta aca att 624 Thr Lys Lys Thr Ala Ala Trp Asn Ser Gly Thr Ser Thr Leu Thr Ile 2gta aac agt aaa aaa act aaa gac ctt gtg ttt aca aaa gaa aac 672 Thr Val Asn Ser Lys Lys Thr Lys Asp Leu Val Phe Thr Lys Glu Asn 222tt aca gta caa caa tac gac tca aat ggc acc aaa tta gag ggg 72le Thr Val Gln Gln Tyr Asp Ser Asn Gly Thr Lys Leu Glu Gly 225 234ca gtt gaa att aca aaa ctt gat gaa att aaa aac gct tta aaa 768 Ser Ala Val Glu Ile Thr Lys Leu Asp Glu Ile Lys Asn Ala Leu Lys 245 25gt cac ccc atg gat gaa aag ctt tta aaa agt aaa gat gat aaa gca 8His Pro Met Asp Glu Lys Leu Leu Lys Ser Lys Asp Asp Lys Ala 267aa gat ggt aaa gcc ttg gat ctt gat cga gaa tta aat tct aaa 864 Ser Lys Asp Gly Lys Ala Leu Asp Leu Asp Arg Glu Leu Asn Ser Lys 275 28ct tct agc aaa gaa aaa agt aaa gcc aag gaa gaa gaa ata acc aag 9Ser Ser Lys Glu Lys Ser Lys Ala Lys Glu Glu Glu Ile Thr Lys 29aag tca cag aaa agc tta ggc gat ttg aat aat gat gaa aat ctt 96ys Ser Gln Lys Ser Leu Gly Asp Leu Asn Asn Asp Glu Asn Leu 33atg atg cca gaa gat caa aaa tta cct gag gtt aaa aaa tta gat agc t Met Pro Glu Asp Gln Lys Leu Pro Glu Val Lys Lys Leu Asp Ser 325 33aa aaa gaa ttt aaa cct gtt tct gag gtt gag aaa tta gat aag att s Lys Glu Phe Lys Pro Val Ser Glu Val Glu Lys Leu Asp Lys Ile 345ag tct aat aac aat gtt gga gaa tta tca ccg tta gat aaa tct e Lys Ser Asn Asn Asn Val Gly Glu Leu Ser Pro Leu Asp Lys Ser 355 36ct tat aaa gac att gat tca aaa gag gag aca gtt aat aaa gat gtt r Tyr Lys Asp Ile Asp Ser Lys Glu Glu Thr Val Asn Lys Asp Val 378tg caa aag act aag cct cag gtt aaa gac caa gtt act tct ttg n Leu Gln Lys Thr Lys Pro Gln Val Lys Asp Gln Val Thr Ser Leu 385 39gaa gat ttg act act atg tct ata gat tcc agt agt cct gta ttt n Glu Asp Leu Thr Thr Met Ser Ile Asp Ser Ser Ser Pro Val Phe 44gag gtt att gat cca att aca aat tta gga act ctt caa ctt att u Glu Val Ile Asp Pro Ile Thr Asn Leu Gly Thr Leu Gln Leu Ile 423ta aat act ggt gtt agg ctt aaa gaa agc act cag caa ggc att p Leu Asn Thr Gly Val Arg Leu Lys Glu Ser Thr Gln Gln Gly Ile 435 44ag cgg tat gga att tat gaa cgt gaa aaa gat ttg gtt gtt att aaa n Arg Tyr Gly Ile Tyr Glu Arg Glu Lys Asp Leu Val Val Ile Lys 456at tca gga aaa gct aag ctt cag ata ctt gat aaa ctt gaa aat t Asp Ser Gly Lys Ala Lys Leu Gln Ile Leu Asp Lys Leu Glu Asn 465 478aa gtg gta tca gag tct aat ttt gag att aat aaa aat tca tct u Lys Val Val Ser Glu Ser Asn Phe Glu Ile Asn Lys Asn Ser Ser 485 49tt tat gtt gat tct aaa atg att tta gta gct gtt agg gat aaa gat u Tyr Val Asp Ser Lys Met Ile Leu Val Ala Val Arg Asp Lys Asp 55agt aat gat tgg aga ttg gcc aaa ttt tct cct aaa aat tta gat r Ser Asn Asp Trp Arg Leu Ala Lys Phe Ser Pro Lys Asn Leu Asp 5525 gag ttt att ctt tca gag aat aaa att atg cct ttt act agc ttt tct u Phe Ile Leu Ser Glu Asn Lys Ile Met Pro Phe Thr Ser Phe Ser 534ga aaa aat ttt att tat ttg caa gat gag ttt aaa agt cta gtt l Arg Lys Asn Phe Ile Tyr Leu Gln Asp Glu Phe Lys Ser Leu Val 545 556ta gat gta aat act tta aaa aaa gtt aag ggt cac c e Leu Asp Val Asn Thr Leu Lys Lys Val Lys Gly His 565 5773 PRT Artificial Sequence Chimeric Protein Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val Asp Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys Asp 2 Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys Gly 35 4r Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys Ala 5 Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln Thr 65 7 Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Lys 85 9l Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Lys Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg Leu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser Lys Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala Ala Lys Lys Thr Ala Ala Trp Asn Ser Gly Thr Ser Thr Leu Thr Ile 2Val Asn Ser Lys Lys Thr Lys Asp Leu Val Phe Thr Lys Glu Asn 222le Thr Val Gln Gln Tyr Asp Ser Asn Gly Thr Lys Leu Glu Gly 225 234la Val Glu Ile Thr Lys Leu Asp Glu Ile Lys Asn Ala Leu Lys 245 25ly His Pro Met Asp Glu Lys Leu Leu Lys Ser Lys Asp Asp Lys Ala 267ys Asp Gly Lys Ala Leu Asp Leu Asp Arg Glu Leu Asn Ser Lys 275 28la Ser Ser Lys Glu Lys Ser Lys Ala Lys Glu Glu Glu Ile Thr Lys 29Lys Ser Gln Lys Ser Leu Gly Asp Leu Asn Asn Asp Glu Asn Leu 33Met Met Pro Glu Asp Gln Lys Leu Pro Glu Val Lys Lys Leu Asp Ser 325 33ys Lys Glu Phe Lys Pro Val Ser Glu Val Glu Lys Leu Asp Lys Ile 345ys Ser Asn Asn Asn Val Gly Glu Leu Ser Pro Leu Asp Lys Ser 355 36er Tyr Lys Asp Ile Asp Ser Lys Glu Glu Thr Val Asn Lys Asp Val 378eu Gln Lys Thr Lys Pro Gln Val Lys Asp Gln Val Thr Ser Leu 385 39Glu Asp Leu Thr Thr Met Ser Ile Asp Ser Ser Ser Pro Val Phe 44Glu Val Ile Asp Pro Ile Thr Asn Leu Gly Thr Leu Gln Leu Ile 423eu Asn Thr Gly Val Arg Leu Lys Glu Ser Thr Gln Gln Gly Ile 435 44ln Arg Tyr Gly Ile Tyr Glu Arg Glu Lys Asp Leu Val Val Ile Lys 456sp Ser Gly Lys Ala Lys Leu Gln Ile Leu Asp Lys Leu Glu Asn 465 478ys Val Val Ser Glu Ser Asn Phe Glu Ile Asn Lys Asn Ser Ser 485 49eu Tyr Val Asp Ser Lys Met Ile Leu Val Ala Val Arg Asp Lys Asp 55Ser Asn Asp Trp Arg Leu Ala Lys Phe Ser Pro Lys Asn Leu Asp 5525 Glu Phe Ile Leu Ser Glu Asn Lys Ile Met Pro Phe Thr Ser Phe Ser 534rg Lys Asn Phe Ile Tyr Leu Gln Asp Glu Phe Lys Ser Leu Val 545 556eu Asp Val Asn Thr Leu Lys Lys Val Lys Gly His 565 57 Artificial Sequence CDS (imeric Nucleic Acid caa aaa ggt gct gag tca att ggt tct caa aaa gaa aat gat cta 48 Ala Gln Lys Gly Ala Glu Ser Ile Gly Ser Gln Lys Glu Asn Asp Leu ctt gaa gac tct agt aaa aaa tca cat caa aac gct aaa caa gac 96 Asn Leu Glu Asp Ser Ser Lys Lys Ser His Gln Asn Ala Lys Gln Asp 2 ctt cct gcg gtg aca gaa gac tca gtg tct ttg ttt aat ggt aat aaa Pro Ala Val Thr Glu Asp Ser Val Ser Leu Phe Asn Gly Asn Lys 35 4t ttt gta agc aaa gaa aaa aat agc tcc ggc aaa tat gat tta aga Phe Val Ser Lys Glu Lys Asn Ser Ser Gly Lys Tyr Asp Leu Arg 5 gca aca att gat cag gtt gaa ctt aaa gga act tcc gat aaa aac aat 24hr Ile Asp Gln Val Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn 65 7 ggt tct gga acc ctt gaa ggt tca aag cct gac aag agt aaa gta aaa 288 Gly Ser Gly Thr Leu Glu Gly Ser Lys Pro Asp Lys Ser Lys Val Lys 85 9a aca gtt tct gct gat tta aac aca gta acc tta gaa gca ttt gat 336 Leu Thr Val Ser Ala Asp Leu Asn Thr Val Thr Leu Glu Ala Phe Asp agc aac caa aaa att tca agt aaa gtt act aaa aaa cag ggg tca 384 Ala Ser Asn Gln Lys Ile Ser Ser Lys Val Thr Lys Lys Gln Gly Ser aca gag gaa act ctc aaa gct aat aaa tta gac tca aag aaa tta 432 Ile Thr Glu Glu Thr Leu Lys Ala Asn Lys Leu Asp Ser Lys Lys Leu aga tca aac gga act aca ctt gaa tac tca caa ata aca gat gct 48rg Ser Asn Gly Thr Thr Leu Glu Tyr Ser Gln Ile Thr Asp Ala gac aat gct aca aaa gca gta gaa act cta aaa aat agc att aag ctt 528 Asp Asn Ala Thr Lys Ala Val Glu Thr Leu Lys Asn Ser Ile Lys Leu gga agt ctt gta gtc gga aaa aca aca gtg gaa att aaa gaa ggt 576 Glu Gly Ser Leu Val Val Gly Lys Thr Thr Val Glu Ile Lys Glu Gly gtt act cta aaa aga gaa att gaa aaa gat gga aaa gta aaa gtc 624 Thr Val Thr Leu Lys Arg Glu Ile Glu Lys Asp Gly Lys Val Lys Val 2ttg aat gac act gca ggt tct aac aaa aaa aca ggt aaa tgg gaa 672 Phe Leu Asn Asp Thr Ala Gly Ser Asn Lys Lys Thr Gly Lys Trp Glu 222gt act agc act tta aca att agt gct gac agc aaa aaa act aaa 72er Thr Ser Thr Leu Thr Ile Ser Ala Asp Ser Lys Lys Thr Lys 225 234tg gtg ttc tta aca gat ggt aca att aca gta caa caa tac aac 768 Asp Leu Val Phe Leu Thr Asp Gly Thr Ile Thr Val Gln Gln Tyr Asn 245 25ca gct gga acc agc cta gaa gga tca gca agt gaa att aaa aat ctt 8Ala Gly Thr Ser Leu Glu Gly Ser Ala Ser Glu Ile Lys Asn Leu 267ag ctt aaa aac gct tta aaa ggt cac ccc atg gct caa tat aac 864 Ser Glu Leu Lys Asn Ala Leu Lys Gly His Pro Met Ala Gln

Tyr Asn 275 28aa atg cac atg tta tca aac aaa tct gct tct caa aat gta aga aca 9Met His Met Leu Ser Asn Lys Ser Ala Ser Gln Asn Val Arg Thr 29gaa gag ctt gga atg cag cct gca aaa att aac aca cca gca tca 96lu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr Pro Ala Ser 33ctt tca ggg ctt caa gcg tct tgg act tta aga gtt cat gtt gga gca u Ser Gly Leu Gln Ala Ser Trp Thr Leu Arg Val His Val Gly Ala 325 33cc caa gat gaa gct att gct gta aat att tat gca gct aat gtt gca r Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ala Ala Asn Val Ala 345tt ttc tct ggt gag gga gct caa act gct cag gct gca ccg gtt n Leu Phe Ser Gly Glu Gly Ala Gln Thr Ala Gln Ala Ala Pro Val 355 36aa gag ggt gtt caa cag gaa gga gct caa cag cca gca cct gct aca n Glu Gly Val Gln Gln Glu Gly Ala Gln Gln Pro Ala Pro Ala Thr 378ct tct caa ggc gga gtt ggt cac c a Pro Ser Gln Gly Gly Val Gly His 385 3993 PRT Artificial Sequence Chimeric Protein Gln Lys Gly Ala Glu Ser Ile Gly Ser Gln Lys Glu Asn Asp Leu Leu Glu Asp Ser Ser Lys Lys Ser His Gln Asn Ala Lys Gln Asp 2 Leu Pro Ala Val Thr Glu Asp Ser Val Ser Leu Phe Asn Gly Asn Lys 35 4e Phe Val Ser Lys Glu Lys Asn Ser Ser Gly Lys Tyr Asp Leu Arg 5 Ala Thr Ile Asp Gln Val Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn 65 7 Gly Ser Gly Thr Leu Glu Gly Ser Lys Pro Asp Lys Ser Lys Val Lys 85 9u Thr Val Ser Ala Asp Leu Asn Thr Val Thr Leu Glu Ala Phe Asp Ser Asn Gln Lys Ile Ser Ser Lys Val Thr Lys Lys Gln Gly Ser Thr Glu Glu Thr Leu Lys Ala Asn Lys Leu Asp Ser Lys Lys Leu Arg Ser Asn Gly Thr Thr Leu Glu Tyr Ser Gln Ile Thr Asp Ala Asp Asn Ala Thr Lys Ala Val Glu Thr Leu Lys Asn Ser Ile Lys Leu Gly Ser Leu Val Val Gly Lys Thr Thr Val Glu Ile Lys Glu Gly Val Thr Leu Lys Arg Glu Ile Glu Lys Asp Gly Lys Val Lys Val 2Leu Asn Asp Thr Ala Gly Ser Asn Lys Lys Thr Gly Lys Trp Glu 222er Thr Ser Thr Leu Thr Ile Ser Ala Asp Ser Lys Lys Thr Lys 225 234eu Val Phe Leu Thr Asp Gly Thr Ile Thr Val Gln Gln Tyr Asn 245 25hr Ala Gly Thr Ser Leu Glu Gly Ser Ala Ser Glu Ile Lys Asn Leu 267lu Leu Lys Asn Ala Leu Lys Gly His Pro Met Ala Gln Tyr Asn 275 28ln Met His Met Leu Ser Asn Lys Ser Ala Ser Gln Asn Val Arg Thr 29Glu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr Pro Ala Ser 33Leu Ser Gly Leu Gln Ala Ser Trp Thr Leu Arg Val His Val Gly Ala 325 33hr Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ala Ala Asn Val Ala 345eu Phe Ser Gly Glu Gly Ala Gln Thr Ala Gln Ala Ala Pro Val 355 36ln Glu Gly Val Gln Gln Glu Gly Ala Gln Gln Pro Ala Pro Ala Thr 378ro Ser Gln Gly Gly Val Gly His 385 39363 DNA Artificial Sequence CDS (362) Chimeric nucleic acid caa aaa ggt gct gag tca att ggt tct caa aaa gaa aat gat cta 48 Ala Gln Lys Gly Ala Glu Ser Ile Gly Ser Gln Lys Glu Asn Asp Leu ctt gaa gac tct agt aaa aaa tca cat caa aac gct aaa caa gac 96 Asn Leu Glu Asp Ser Ser Lys Lys Ser His Gln Asn Ala Lys Gln Asp 2 ctt cct gcg gtg aca gaa gac tca gtg tct ttg ttt aat ggt aat aaa Pro Ala Val Thr Glu Asp Ser Val Ser Leu Phe Asn Gly Asn Lys 35 4t ttt gta agc aaa gaa aaa aat agc tcc ggc aaa tat gat tta aga Phe Val Ser Lys Glu Lys Asn Ser Ser Gly Lys Tyr Asp Leu Arg 5 gca aca att gat cag gtt gaa ctt aaa gga act tcc gat aaa aac aat 24hr Ile Asp Gln Val Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn 65 7 ggt tct gga acc ctt gaa ggt tca aag cct gac aag agt aaa gta aaa 288 Gly Ser Gly Thr Leu Glu Gly Ser Lys Pro Asp Lys Ser Lys Val Lys 85 9a aca gtt tct gct gat tta aac aca gta acc tta gaa gca ttt gat 336 Leu Thr Val Ser Ala Asp Leu Asn Thr Val Thr Leu Glu Ala Phe Asp agc aac caa aaa att tca agt aaa gtt act aaa aaa cag ggg tca 384 Ala Ser Asn Gln Lys Ile Ser Ser Lys Val Thr Lys Lys Gln Gly Ser aca gag gaa act ctc aaa gct aat aaa tta gac tca aag aaa tta 432 Ile Thr Glu Glu Thr Leu Lys Ala Asn Lys Leu Asp Ser Lys Lys Leu aga tca aac gga act aca ctt gaa tac tca caa ata aca gat gct 48rg Ser Asn Gly Thr Thr Leu Glu Tyr Ser Gln Ile Thr Asp Ala gac aat gct aca aaa gca gta gaa act cta aaa aat agc att aag ctt 528 Asp Asn Ala Thr Lys Ala Val Glu Thr Leu Lys Asn Ser Ile Lys Leu gga agt ctt gta gtc gga aaa aca aca gtg gaa att aaa gaa ggt 576 Glu Gly Ser Leu Val Val Gly Lys Thr Thr Val Glu Ile Lys Glu Gly gtt act cta aaa aga gaa att gaa aaa gat gga aaa gta aaa gtc 624 Thr Val Thr Leu Lys Arg Glu Ile Glu Lys Asp Gly Lys Val Lys Val 2ttg aat gac act gca ggt tct aac aaa aaa aca ggt aaa tgg gaa 672 Phe Leu Asn Asp Thr Ala Gly Ser Asn Lys Lys Thr Gly Lys Trp Glu 222gt act agc act tta aca att agt gct gac agc aaa aaa act aaa 72er Thr Ser Thr Leu Thr Ile Ser Ala Asp Ser Lys Lys Thr Lys 225 234tg gtg ttc tta aca gat ggt aca att aca gta caa caa tac aac 768 Asp Leu Val Phe Leu Thr Asp Gly Thr Ile Thr Val Gln Gln Tyr Asn 245 25ca gct gga acc agc cta gaa gga tca gca agt gaa att aaa aat ctt 8Ala Gly Thr Ser Leu Glu Gly Ser Ala Ser Glu Ile Lys Asn Leu 267ag ctt aaa aac gct tta aaa ggt cac ccc atg gct caa tat aac 864 Ser Glu Leu Lys Asn Ala Leu Lys Gly His Pro Met Ala Gln Tyr Asn 275 28aa atg cac atg tta tca aac aaa tct gct tct caa aat gta aga aca 9Met His Met Leu Ser Asn Lys Ser Ala Ser Gln Asn Val Arg Thr 29gaa gag ctt gga atg cag cct gca aaa att aac aca cca gca tca 96lu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr Pro Ala Ser 33ctt tca ggg ctt caa gcg tct tgg act tta aga gtt cat gtt gga gca u Ser Gly Leu Gln Ala Ser Trp Thr Leu Arg Val His Val Gly Ala 325 33cc caa gat gaa gct att gct gta aat att tat gca gct aat gtt gca r Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ala Ala Asn Val Ala 345tt ttc tct ggt gag gga gct caa act gct cag gct gca ccg gtt n Leu Phe Ser Gly Glu Gly Ala Gln Thr Ala Gln Ala Ala Pro Val 355 36aa gag ggt gtt caa cag gaa gga gct caa cag cca gca cct gct aca n Glu Gly Val Gln Gln Glu Gly Ala Gln Gln Pro Ala Pro Ala Thr 378ct tct caa ggc gga gtt aat tct cct gtt aat gtt aca act aca a Pro Ser Gln Gly Gly Val Asn Ser Pro Val Asn Val Thr Thr Thr 385 39gat gct aat aca tca ctt gct aaa att gaa aat gct att aga atg l Asp Ala Asn Thr Ser Leu Ala Lys Ile Glu Asn Ala Ile Arg Met 44agt gat caa agg gca aat tta ggt gct ttc caa aat aga ctt gaa e Ser Asp Gln Arg Ala Asn Leu Gly Ala Phe Gln Asn Arg Leu Glu 423ta aag aat agt act gag tat gca att gaa aat cta aaa gca tct r Ile Lys Asn Ser Thr Glu Tyr Ala Ile Glu Asn Leu Lys Ala Ser 435 44at gct caa ata ggt cac c r Ala Gln Ile Gly His 4554 PRT Artificial Sequence Chimeric Protein Gln Lys Gly Ala Glu Ser Ile Gly Ser Gln Lys Glu Asn Asp Leu Leu Glu Asp Ser Ser Lys Lys Ser His Gln Asn Ala Lys Gln Asp 2 Leu Pro Ala Val Thr Glu Asp Ser Val Ser Leu Phe Asn Gly Asn Lys 35 4e Phe Val Ser Lys Glu Lys Asn Ser Ser Gly Lys Tyr Asp Leu Arg 5 Ala Thr Ile Asp Gln Val Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn 65 7 Gly Ser Gly Thr Leu Glu Gly Ser Lys Pro Asp Lys Ser Lys Val Lys 85 9u Thr Val Ser Ala Asp Leu Asn Thr Val Thr Leu Glu Ala Phe Asp Ser Asn Gln Lys Ile Ser Ser Lys Val Thr Lys Lys Gln Gly Ser Thr Glu Glu Thr Leu Lys Ala Asn Lys Leu Asp Ser Lys Lys Leu Arg Ser Asn Gly Thr Thr Leu Glu Tyr Ser Gln Ile Thr Asp Ala Asp Asn Ala Thr Lys Ala Val Glu Thr Leu Lys Asn Ser Ile Lys Leu Gly Ser Leu Val Val Gly Lys Thr Thr Val Glu Ile Lys Glu Gly Val Thr Leu Lys Arg Glu Ile Glu Lys Asp Gly Lys Val Lys Val 2Leu Asn Asp Thr Ala Gly Ser Asn Lys Lys Thr Gly Lys Trp Glu 222er Thr Ser Thr Leu Thr Ile Ser Ala Asp Ser Lys Lys Thr Lys 225 234eu Val Phe Leu Thr Asp Gly Thr Ile Thr Val Gln Gln Tyr Asn 245 25hr Ala Gly Thr Ser Leu Glu Gly Ser Ala Ser Glu Ile Lys Asn Leu 267lu Leu Lys Asn Ala Leu Lys Gly His Pro Met Ala Gln Tyr Asn 275 28ln Met His Met Leu Ser Asn Lys Ser Ala Ser Gln Asn Val Arg Thr 29Glu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr Pro Ala Ser 33Leu Ser Gly Leu Gln Ala Ser Trp Thr Leu Arg Val His Val Gly Ala 325 33hr Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ala Ala Asn Val Ala 345eu Phe Ser Gly Glu Gly Ala Gln Thr Ala Gln Ala Ala Pro Val 355 36ln Glu Gly Val Gln Gln Glu Gly Ala Gln Gln Pro Ala Pro Ala Thr 378ro Ser Gln Gly Gly Val Asn Ser Pro Val Asn Val Thr Thr Thr 385 39Asp Ala Asn Thr Ser Leu Ala Lys Ile Glu Asn Ala Ile Arg Met 44Ser Asp Gln Arg Ala Asn Leu Gly Ala Phe Gln Asn Arg Leu Glu 423le Lys Asn Ser Thr Glu Tyr Ala Ile Glu Asn Leu Lys Ala Ser 435 44yr Ala Gln Ile Gly His 45 Artificial Sequence CDS (imeric nucleic acid caa aaa ggt gct gag tca att ggt tct caa aaa gaa aat gat cta 48 Ala Gln Lys Gly Ala Glu Ser Ile Gly Ser Gln Lys Glu Asn Asp Leu ctt gaa gac tct agt aaa aaa tca cat caa aac gct aaa caa gac 96 Asn Leu Glu Asp Ser Ser Lys Lys Ser His Gln Asn Ala Lys Gln Asp 2 ctt cct gcg gtg aca gaa gac tca gtg tct ttg ttt aat ggt aat aaa Pro Ala Val Thr Glu Asp Ser Val Ser Leu Phe Asn Gly Asn Lys 35 4t ttt gta agc aaa gaa aaa aat agc tcc ggc aaa tat gat tta aga Phe Val Ser Lys Glu Lys Asn Ser Ser Gly Lys Tyr Asp Leu Arg 5 gca aca att gat cag gtt gaa ctt aaa gga act tcc gat aaa aac aat 24hr Ile Asp Gln Val Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn 65 7 ggt tct gga acc ctt gaa ggt tca aag cct gac aag agt aaa gta aaa 288 Gly Ser Gly Thr Leu Glu Gly Ser Lys Pro Asp Lys Ser Lys Val Lys 85 9a aca gtt tct gct gat tta aac aca gta acc tta gaa gca ttt gat 336 Leu Thr Val Ser Ala Asp Leu Asn Thr Val Thr Leu Glu Ala Phe Asp agc aac caa aaa att tca agt aaa gtt act aaa aaa cag ggg tca 384 Ala Ser Asn Gln Lys Ile Ser Ser Lys Val Thr Lys Lys Gln Gly Ser aca gag gaa act ctc aaa gct aat aaa tta gac tca aag aaa tta 432 Ile Thr Glu Glu Thr Leu Lys Ala Asn Lys Leu Asp Ser Lys Lys Leu aga tca aac gga act aca ctt gaa tac tca caa ata aca gat gct 48rg Ser Asn Gly Thr Thr Leu Glu Tyr Ser Gln Ile Thr Asp Ala gac aat gct aca aaa gca gta gaa act cta aaa aat agc att aag ctt 528 Asp Asn Ala Thr Lys Ala Val Glu Thr Leu Lys Asn Ser Ile Lys Leu gga agt ctt gta gtc gga aaa aca aca gtg gaa att aaa gaa ggt 576 Glu Gly Ser Leu Val Val Gly Lys Thr Thr Val Glu Ile Lys Glu Gly gtt act cta aaa aga gaa att gaa aaa gat gga aaa gta aaa gtc 624 Thr Val Thr Leu Lys Arg Glu Ile Glu Lys Asp Gly Lys Val Lys Val 2ttg aat gac act gca ggt tct aac aaa aaa aca ggt aaa tgg gaa 672 Phe Leu Asn Asp Thr Ala Gly Ser Asn Lys Lys Thr Gly Lys Trp Glu 222gt act agc act tta aca att agt gct gac agc aaa aaa act aaa 72er Thr Ser Thr Leu Thr Ile Ser Ala Asp Ser Lys Lys Thr Lys 225 234tg gtg ttc tta aca gat ggt aca att aca gta caa caa tac aac 768 Asp Leu Val Phe Leu Thr Asp Gly Thr Ile Thr Val Gln Gln Tyr Asn 245 25ca gct gga acc agc cta gaa gga tca gca agt gaa att aaa aat ctt 8Ala Gly Thr Ser Leu Glu Gly Ser Ala Ser Glu Ile Lys Asn Leu 267ag ctt aaa aac gct tta aaa ggt cac ccc atg gct tct caa aat 864 Ser Glu Leu Lys Asn Ala Leu Lys Gly His Pro Met Ala Ser Gln Asn 275 28ta aga aca gct gaa gag ctt gga atg cag cct gca aaa att aac aca 9Arg Thr Ala Glu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr 29gca tca ctt tca ggg ctt caa gcg tct tgg act tta aga gtt cat 96la Ser Leu Ser Gly Leu Gln Ala Ser Trp Thr Leu Arg Val His 33gtt gga gca acc caa gat gaa gct att gct gta aat att tat gca gct l Gly Ala Thr Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ala Ala 325 33at gtt gca aat ctt ttc tct ggt gag gga gct caa act gct cag gct n Val Ala Asn Leu Phe Ser Gly Glu Gly Ala Gln Thr Ala Gln Ala 345cg gtt caa gag ggt gtt caa cag gaa gga gct caa cag cca gca a Pro Val Gln Glu Gly Val Gln Gln Glu Gly Ala Gln Gln Pro Ala 355 36ct gct aca gca cct tct caa ggc gga gtt ggt cac c o Ala Thr Ala Pro Ser Gln Gly Gly Val Gly His 3788rtificial Sequence Chimeric Protein Gln Lys Gly Ala Glu Ser Ile Gly Ser Gln Lys Glu Asn Asp Leu Leu Glu Asp Ser Ser Lys Lys Ser His Gln Asn Ala Lys Gln Asp 2 Leu Pro Ala Val Thr Glu Asp Ser Val Ser Leu Phe

Asn Gly Asn Lys 35 4e Phe Val Ser Lys Glu Lys Asn Ser Ser Gly Lys Tyr Asp Leu Arg 5 Ala Thr Ile Asp Gln Val Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn 65 7 Gly Ser Gly Thr Leu Glu Gly Ser Lys Pro Asp Lys Ser Lys Val Lys 85 9u Thr Val Ser Ala Asp Leu Asn Thr Val Thr Leu Glu Ala Phe Asp Ser Asn Gln Lys Ile Ser Ser Lys Val Thr Lys Lys Gln Gly Ser Thr Glu Glu Thr Leu Lys Ala Asn Lys Leu Asp Ser Lys Lys Leu Arg Ser Asn Gly Thr Thr Leu Glu Tyr Ser Gln Ile Thr Asp Ala Asp Asn Ala Thr Lys Ala Val Glu Thr Leu Lys Asn Ser Ile Lys Leu Gly Ser Leu Val Val Gly Lys Thr Thr Val Glu Ile Lys Glu Gly Val Thr Leu Lys Arg Glu Ile Glu Lys Asp Gly Lys Val Lys Val 2Leu Asn Asp Thr Ala Gly Ser Asn Lys Lys Thr Gly Lys Trp Glu 222er Thr Ser Thr Leu Thr Ile Ser Ala Asp Ser Lys Lys Thr Lys 225 234eu Val Phe Leu Thr Asp Gly Thr Ile Thr Val Gln Gln Tyr Asn 245 25hr Ala Gly Thr Ser Leu Glu Gly Ser Ala Ser Glu Ile Lys Asn Leu 267lu Leu Lys Asn Ala Leu Lys Gly His Pro Met Ala Ser Gln Asn 275 28al Arg Thr Ala Glu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr 29Ala Ser Leu Ser Gly Leu Gln Ala Ser Trp Thr Leu Arg Val His 33Val Gly Ala Thr Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ala Ala 325 33sn Val Ala Asn Leu Phe Ser Gly Glu Gly Ala Gln Thr Ala Gln Ala 345ro Val Gln Glu Gly Val Gln Gln Glu Gly Ala Gln Gln Pro Ala 355 36ro Ala Thr Ala Pro Ser Gln Gly Gly Val Gly His 378324 DNA Artificial Sequence CDS (323) Chimeric nucleic acid caa aaa ggt gct gag tca att ggt tct caa aaa gaa aat gat cta 48 Ala Gln Lys Gly Ala Glu Ser Ile Gly Ser Gln Lys Glu Asn Asp Leu ctt gaa gac tct agt aaa aaa tca cat caa aac gct aaa caa gac 96 Asn Leu Glu Asp Ser Ser Lys Lys Ser His Gln Asn Ala Lys Gln Asp 2 ctt cct gcg gtg aca gaa gac tca gtg tct ttg ttt aat ggt aat aaa Pro Ala Val Thr Glu Asp Ser Val Ser Leu Phe Asn Gly Asn Lys 35 4t ttt gta agc aaa gaa aaa aat agc tcc ggc aaa tat gat tta aga Phe Val Ser Lys Glu Lys Asn Ser Ser Gly Lys Tyr Asp Leu Arg 5 gca aca att gat cag gtt gaa ctt aaa gga act tcc gat aaa aac aat 24hr Ile Asp Gln Val Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn 65 7 ggt tct gga acc ctt gaa ggt tca aag cct gac aag agt aaa gta aaa 288 Gly Ser Gly Thr Leu Glu Gly Ser Lys Pro Asp Lys Ser Lys Val Lys 85 9a aca gtt tct gct gat tta aac aca gta acc tta gaa gca ttt gat 336 Leu Thr Val Ser Ala Asp Leu Asn Thr Val Thr Leu Glu Ala Phe Asp agc aac caa aaa att tca agt aaa gtt act aaa aaa cag ggg tca 384 Ala Ser Asn Gln Lys Ile Ser Ser Lys Val Thr Lys Lys Gln Gly Ser aca gag gaa act ctc aaa gct aat aaa tta gac tca aag aaa tta 432 Ile Thr Glu Glu Thr Leu Lys Ala Asn Lys Leu Asp Ser Lys Lys Leu aga tca aac gga act aca ctt gaa tac tca caa ata aca gat gct 48rg Ser Asn Gly Thr Thr Leu Glu Tyr Ser Gln Ile Thr Asp Ala gac aat gct aca aaa gca gta gaa act cta aaa aat agc att aag ctt 528 Asp Asn Ala Thr Lys Ala Val Glu Thr Leu Lys Asn Ser Ile Lys Leu gga agt ctt gta gtc gga aaa aca aca gtg gaa att aaa gaa ggt 576 Glu Gly Ser Leu Val Val Gly Lys Thr Thr Val Glu Ile Lys Glu Gly gtt act cta aaa aga gaa att gaa aaa gat gga aaa gta aaa gtc 624 Thr Val Thr Leu Lys Arg Glu Ile Glu Lys Asp Gly Lys Val Lys Val 2ttg aat gac act gca ggt tct aac aaa aaa aca ggt aaa tgg gaa 672 Phe Leu Asn Asp Thr Ala Gly Ser Asn Lys Lys Thr Gly Lys Trp Glu 222gt act agc act tta aca att agt gct gac agc aaa aaa act aaa 72er Thr Ser Thr Leu Thr Ile Ser Ala Asp Ser Lys Lys Thr Lys 225 234tg gtg ttc tta aca gat ggt aca att aca gta caa caa tac aac 768 Asp Leu Val Phe Leu Thr Asp Gly Thr Ile Thr Val Gln Gln Tyr Asn 245 25ca gct gga acc agc cta gaa gga tca gca agt gaa att aaa aat ctt 8Ala Gly Thr Ser Leu Glu Gly Ser Ala Ser Glu Ile Lys Asn Leu 267ag ctt aaa aac gct tta aaa ggt cac ccc atg gct tct caa aat 864 Ser Glu Leu Lys Asn Ala Leu Lys Gly His Pro Met Ala Ser Gln Asn 275 28ta aga aca gct gaa gag ctt gga atg cag cct gca aaa att aac aca 9Arg Thr Ala Glu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr 29gca tca ctt tca ggg ctt caa gcg tct tgg act tta aga gtt cat 96la Ser Leu Ser Gly Leu Gln Ala Ser Trp Thr Leu Arg Val His 33gtt gga gca acc caa gat gaa gct att gct gta aat att tat gca gct l Gly Ala Thr Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ala Ala 325 33at gtt gca aat ctt ttc tct ggt gag gga gct caa act gct cag gct n Val Ala Asn Leu Phe Ser Gly Glu Gly Ala Gln Thr Ala Gln Ala 345cg gtt caa gag ggt gtt caa cag gaa gga gct caa cag cca gca a Pro Val Gln Glu Gly Val Gln Gln Glu Gly Ala Gln Gln Pro Ala 355 36ct gct aca gca cct tct caa ggc gga gtt aat tct cct gtt aat gtt o Ala Thr Ala Pro Ser Gln Gly Gly Val Asn Ser Pro Val Asn Val 378ct aca gtt gat gct aat aca tca ctt gct aaa att gaa aat gct r Thr Thr Val Asp Ala Asn Thr Ser Leu Ala Lys Ile Glu Asn Ala 385 39aga atg ata agt gat caa agg gca aat tta ggt gct ttc caa aat e Arg Met Ile Ser Asp Gln Arg Ala Asn Leu Gly Ala Phe Gln Asn 44ctt gaa tct ata aag aat agt act gag tat gca att gaa aat cta g Leu Glu Ser Ile Lys Asn Ser Thr Glu Tyr Ala Ile Glu Asn Leu 423ca tct tat gct caa ata ggt cac c s Ala Ser Tyr Ala Gln Ile Gly His 435 444rtificial Sequence Chimeric Protein Gln Lys Gly Ala Glu Ser Ile Gly Ser Gln Lys Glu Asn Asp Leu Leu Glu Asp Ser Ser Lys Lys Ser His Gln Asn Ala Lys Gln Asp 2 Leu Pro Ala Val Thr Glu Asp Ser Val Ser Leu Phe Asn Gly Asn Lys 35 4e Phe Val Ser Lys Glu Lys Asn Ser Ser Gly Lys Tyr Asp Leu Arg 5 Ala Thr Ile Asp Gln Val Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn 65 7 Gly Ser Gly Thr Leu Glu Gly Ser Lys Pro Asp Lys Ser Lys Val Lys 85 9u Thr Val Ser Ala Asp Leu Asn Thr Val Thr Leu Glu Ala Phe Asp Ser Asn Gln Lys Ile Ser Ser Lys Val Thr Lys Lys Gln Gly Ser Thr Glu Glu Thr Leu Lys Ala Asn Lys Leu Asp Ser Lys Lys Leu Arg Ser Asn Gly Thr Thr Leu Glu Tyr Ser Gln Ile Thr Asp Ala Asp Asn Ala Thr Lys Ala Val Glu Thr Leu Lys Asn Ser Ile Lys Leu Gly Ser Leu Val Val Gly Lys Thr Thr Val Glu Ile Lys Glu Gly Val Thr Leu Lys Arg Glu Ile Glu Lys Asp Gly Lys Val Lys Val 2Leu Asn Asp Thr Ala Gly Ser Asn Lys Lys Thr Gly Lys Trp Glu 222er Thr Ser Thr Leu Thr Ile Ser Ala Asp Ser Lys Lys Thr Lys 225 234eu Val Phe Leu Thr Asp Gly Thr Ile Thr Val Gln Gln Tyr Asn 245 25hr Ala Gly Thr Ser Leu Glu Gly Ser Ala Ser Glu Ile Lys Asn Leu 267lu Leu Lys Asn Ala Leu Lys Gly His Pro Met Ala Ser Gln Asn 275 28al Arg Thr Ala Glu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr 29Ala Ser Leu Ser Gly Leu Gln Ala Ser Trp Thr Leu Arg Val His 33Val Gly Ala Thr Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ala Ala 325 33sn Val Ala Asn Leu Phe Ser Gly Glu Gly Ala Gln Thr Ala Gln Ala 345ro Val Gln Glu Gly Val Gln Gln Glu Gly Ala Gln Gln Pro Ala 355 36ro Ala Thr Ala Pro Ser Gln Gly Gly Val Asn Ser Pro Val Asn Val 378hr Thr Val Asp Ala Asn Thr Ser Leu Ala Lys Ile Glu Asn Ala 385 39Arg Met Ile Ser Asp Gln Arg Ala Asn Leu Gly Ala Phe Gln Asn 44Leu Glu Ser Ile Lys Asn Ser Thr Glu Tyr Ala Ile Glu Asn Leu 423la Ser Tyr Ala Gln Ile Gly His 435 44765 DNA Artificial Sequence CDS (764) Chimeric nucleic acid caa aaa ggt gct gag tca att ggt tct caa aaa gaa aat gat cta 48 Ala Gln Lys Gly Ala Glu Ser Ile Gly Ser Gln Lys Glu Asn Asp Leu ctt gaa gac tct agt aaa aaa tca cat caa aac gct aaa caa gac 96 Asn Leu Glu Asp Ser Ser Lys Lys Ser His Gln Asn Ala Lys Gln Asp 2 ctt cct gcg gtg aca gaa gac tca gtg tct ttg ttt aat ggt aat aaa Pro Ala Val Thr Glu Asp Ser Val Ser Leu Phe Asn Gly Asn Lys 35 4t ttt gta agc aaa gaa aaa aat agc tcc ggc aaa tat gat tta aga Phe Val Ser Lys Glu Lys Asn Ser Ser Gly Lys Tyr Asp Leu Arg 5 gca aca att gat cag gtt gaa ctt aaa gga act tcc gat aaa aac aat 24hr Ile Asp Gln Val Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn 65 7 ggt tct gga acc ctt gaa ggt tca aag cct gac aag agt aaa gta aaa 288 Gly Ser Gly Thr Leu Glu Gly Ser Lys Pro Asp Lys Ser Lys Val Lys 85 9a aca gtt tct gct gat tta aac aca gta acc tta gaa gca ttt gat 336 Leu Thr Val Ser Ala Asp Leu Asn Thr Val Thr Leu Glu Ala Phe Asp agc aac caa aaa att tca agt aaa gtt act aaa aaa cag ggg tca 384 Ala Ser Asn Gln Lys Ile Ser Ser Lys Val Thr Lys Lys Gln Gly Ser aca gag gaa act ctc aaa gct aat aaa tta gac tca aag aaa tta 432 Ile Thr Glu Glu Thr Leu Lys Ala Asn Lys Leu Asp Ser Lys Lys Leu aga tca aac gga act aca ctt gaa tac tca caa ata aca gat gct 48rg Ser Asn Gly Thr Thr Leu Glu Tyr Ser Gln Ile Thr Asp Ala gac aat gct aca aaa gca gta gaa act cta aaa aat agc att aag ctt 528 Asp Asn Ala Thr Lys Ala Val Glu Thr Leu Lys Asn Ser Ile Lys Leu gga agt ctt gta gtc gga aaa aca aca gtg gaa att aaa gaa ggt 576 Glu Gly Ser Leu Val Val Gly Lys Thr Thr Val Glu Ile Lys Glu Gly gtt act cta aaa aga gaa att gaa aaa gat gga aaa gta aaa gtc 624 Thr Val Thr Leu Lys Arg Glu Ile Glu Lys Asp Gly Lys Val Lys Val 2ttg aat gac act gca ggt tct aac aaa aaa aca ggt aaa tgg gaa 672 Phe Leu Asn Asp Thr Ala Gly Ser Asn Lys Lys Thr Gly Lys Trp Glu 222gt act agc act tta aca att agt gct gac agc aaa aaa act aaa 72er Thr Ser Thr Leu Thr Ile Ser Ala Asp Ser Lys Lys Thr Lys 225 234tg gtg ttc tta aca gat ggt aca att aca gta caa caa tac aac 768 Asp Leu Val Phe Leu Thr Asp Gly Thr Ile Thr Val Gln Gln Tyr Asn 245 25ca gct gga acc agc cta gaa gga tca gca agt gaa att aaa aat ctt 8Ala Gly Thr Ser Leu Glu Gly Ser Ala Ser Glu Ile Lys Asn Leu 267ag ctt aaa aac gct tta aaa ggt cac ccc atg gga aat aat tca 864 Ser Glu Leu Lys Asn Ala Leu Lys Gly His Pro Met Gly Asn Asn Ser 275 28gg aaa gat ggg aat aca tct gca aat tct gct gat gag tct gtt aaa 9Lys Asp Gly Asn Thr Ser Ala Asn Ser Ala Asp Glu Ser Val Lys 29cct aat ctt aca gaa ata agt aaa aaa att acg gat tct aat gcg 96ro Asn Leu Thr Glu Ile Ser Lys Lys Ile Thr Asp Ser Asn Ala 33gtt tta ctt gct gtg aaa gag gtt gaa gcg ttg ctg tca tct ata gat l Leu Leu Ala Val Lys Glu Val Glu Ala Leu Leu Ser Ser Ile Asp 325 33aa att gct gct aaa gct att ggt aaa aaa ata cac caa aat aat ggt u Ile Ala Ala Lys Ala Ile Gly Lys Lys Ile His Gln Asn Asn Gly 345at acc gaa tat aat cac aat gga tca ttg tta gcg gga cgt tat u Asp Thr Glu Tyr Asn His Asn Gly Ser Leu Leu Ala Gly Arg Tyr 355 36ca ata tca acc cta ata aaa caa aaa tta gat gga ttg aaa aat gaa a Ile Ser Thr Leu Ile Lys Gln Lys Leu Asp Gly Leu Lys Asn Glu 378ta aag gaa aaa att gat gcg gct aag aaa tgt tct gaa aca ttt y Leu Lys Glu Lys Ile Asp Ala Ala Lys Lys Cys Ser Glu Thr Phe 385 39aat aaa tta aaa gaa aaa cac aca gat ctt ggt aaa gaa ggt gtt r Asn Lys Leu Lys Glu Lys His Thr Asp Leu Gly Lys Glu Gly Val 44gat gct gat gca aaa gaa gcc att tta aaa aca aat ggt act aaa r Asp Ala Asp Ala Lys Glu Ala Ile Leu Lys Thr Asn Gly Thr Lys 423aa ggt gct gaa gaa ctt gga aaa tta ttt gaa tca gta gag gtc r Lys Gly Ala Glu Glu Leu Gly Lys Leu Phe Glu Ser Val Glu Val 435 44tg tca aaa gca gct aaa gag atg ctt gct aat tca gtt aaa gag ctt u Ser Lys Ala Ala Lys Glu Met Leu Ala Asn Ser Val Lys Glu Leu 456gc cct gtt gtg gca gaa agt cca aaa aaa cct ggt acc atg gct r Ser Pro Val Val Ala Glu Ser Pro Lys Lys Pro Gly Thr Met Ala 465 478at aac caa atg cac atg tta tca aac aaa tct gct tct caa aat n Tyr Asn Gln Met His Met Leu Ser Asn Lys Ser Ala Ser Gln Asn 485 49ta aga aca gct gaa gag ctt gga atg cag cct gca aaa att aac aca l Arg Thr Ala Glu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr 55gca tca ctt tca ggg ctt caa gcg tct tgg act tta aga gtt cat o Ala Ser Leu Ser Gly Leu Gln Ala Ser Trp Thr Leu Arg Val His 5525 gtt gga gca acc caa gat gaa gct att gct gta aat att tat gca gct l Gly Ala Thr Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ala Ala 534tt gca aat ctt ttc tct ggt gag gga gct caa act gct cag gct n Val Ala Asn Leu Phe Ser Gly Glu Gly Ala Gln Thr Ala Gln Ala 545 556cg gtt caa gag ggt gtt caa cag gaa gga gct caa cag cca gca a Pro Val Gln Glu Gly Val Gln Gln Glu Gly Ala Gln Gln Pro Ala 565 57ct

gct aca gca cct tct caa ggc gga gtt ggt cac c o Ala Thr Ala Pro Ser Gln Gly Gly Val Gly His 5822 588 PRT Artificial Sequence Chimeric Protein Gln Lys Gly Ala Glu Ser Ile Gly Ser Gln Lys Glu Asn Asp Leu Leu Glu Asp Ser Ser Lys Lys Ser His Gln Asn Ala Lys Gln Asp 2 Leu Pro Ala Val Thr Glu Asp Ser Val Ser Leu Phe Asn Gly Asn Lys 35 4e Phe Val Ser Lys Glu Lys Asn Ser Ser Gly Lys Tyr Asp Leu Arg 5 Ala Thr Ile Asp Gln Val Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn 65 7 Gly Ser Gly Thr Leu Glu Gly Ser Lys Pro Asp Lys Ser Lys Val Lys 85 9u Thr Val Ser Ala Asp Leu Asn Thr Val Thr Leu Glu Ala Phe Asp Ser Asn Gln Lys Ile Ser Ser Lys Val Thr Lys Lys Gln Gly Ser Thr Glu Glu Thr Leu Lys Ala Asn Lys Leu Asp Ser Lys Lys Leu Arg Ser Asn Gly Thr Thr Leu Glu Tyr Ser Gln Ile Thr Asp Ala Asp Asn Ala Thr Lys Ala Val Glu Thr Leu Lys Asn Ser Ile Lys Leu Gly Ser Leu Val Val Gly Lys Thr Thr Val Glu Ile Lys Glu Gly Val Thr Leu Lys Arg Glu Ile Glu Lys Asp Gly Lys Val Lys Val 2Leu Asn Asp Thr Ala Gly Ser Asn Lys Lys Thr Gly Lys Trp Glu 222er Thr Ser Thr Leu Thr Ile Ser Ala Asp Ser Lys Lys Thr Lys 225 234eu Val Phe Leu Thr Asp Gly Thr Ile Thr Val Gln Gln Tyr Asn 245 25hr Ala Gly Thr Ser Leu Glu Gly Ser Ala Ser Glu Ile Lys Asn Leu 267lu Leu Lys Asn Ala Leu Lys Gly His Pro Met Gly Asn Asn Ser 275 28ly Lys Asp Gly Asn Thr Ser Ala Asn Ser Ala Asp Glu Ser Val Lys 29Pro Asn Leu Thr Glu Ile Ser Lys Lys Ile Thr Asp Ser Asn Ala 33Val Leu Leu Ala Val Lys Glu Val Glu Ala Leu Leu Ser Ser Ile Asp 325 33lu Ile Ala Ala Lys Ala Ile Gly Lys Lys Ile His Gln Asn Asn Gly 345sp Thr Glu Tyr Asn His Asn Gly Ser Leu Leu Ala Gly Arg Tyr 355 36la Ile Ser Thr Leu Ile Lys Gln Lys Leu Asp Gly Leu Lys Asn Glu 378eu Lys Glu Lys Ile Asp Ala Ala Lys Lys Cys Ser Glu Thr Phe 385 39Asn Lys Leu Lys Glu Lys His Thr Asp Leu Gly Lys Glu Gly Val 44Asp Ala Asp Ala Lys Glu Ala Ile Leu Lys Thr Asn Gly Thr Lys 423ys Gly Ala Glu Glu Leu Gly Lys Leu Phe Glu Ser Val Glu Val 435 44eu Ser Lys Ala Ala Lys Glu Met Leu Ala Asn Ser Val Lys Glu Leu 456er Pro Val Val Ala Glu Ser Pro Lys Lys Pro Gly Thr Met Ala 465 478yr Asn Gln Met His Met Leu Ser Asn Lys Ser Ala Ser Gln Asn 485 49al Arg Thr Ala Glu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr 55Ala Ser Leu Ser Gly Leu Gln Ala Ser Trp Thr Leu Arg Val His 5525 Val Gly Ala Thr Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ala Ala 534al Ala Asn Leu Phe Ser Gly Glu Gly Ala Gln Thr Ala Gln Ala 545 556ro Val Gln Glu Gly Val Gln Gln Glu Gly Ala Gln Gln Pro Ala 565 57ro Ala Thr Ala Pro Ser Gln Gly Gly Val Gly His 5823 7Borrelia burgdorferi ctgttaa gtttattttt attgctctca atatcttgtt ctttagataa tgaaggtgta 6aaaag attacgagtc aaaaaaacag agtatactag gtgaattaaa tcagctattg caaacta caaattcact aaaagaagca aaaaatacaa cagataattt aaatgcatca gaggcaa ataaagttgt agaagcagtt ataagtgtgg ttaatttaat ttcatctgct 24tcagg taaaaggtca acaacaaata tgcacgattt agctcaaatg gcagaaatag 3agaaaa aataaaggaa tctagtgata aagtaatagt tgcggctaat gttgcgaaag 36tataa ccttactaaa gcagtagaac aaaatatgca aaaactgtac aaagagcaag 42caact aaaacactat ctgattctga tgaaacagaa cgagtttctg atgaaataaa 48ctaaa gaggctgtag aaatagcttg gaaagccaca gtaaaagtaa aagatgagtt 54atgta gaaaatgcag tcaaagaggc attggataaa ataaagacag aaaccgcgaa 6acaaaa cttacagata tagaagaagt agcagagtta gtattacaga tagccaaaaa 66cggaa atagcgcaag aagttgtggc cttgttaaat actt 77Borrelia burgdorferi ctgttaa gtttattttt attgctctca atatcttgtt atttagataa tgaaggtgta 6aaaag attacgagtc aaaaaaacag agtatactag gtgaattaaa tcagctattg caaacta caaattcact aaaagaagca aaaaatacaa cagataattt aaatgcatca gaggcaa ataaagttgt agaagcagtt ataagtgtgg ttaatttaat ttcatctgct 24tcagg taaaaggtca acaacaaata tgcacgattt agctcaaatg gcagaaatag 3agaaaa aataaaggaa tctagtgata aagtaatagt tgcggctaat gttgcgaaag 36tataa ccttactaaa gcagtagaac aaaatatgca aaaactgtac aaagagcaag 42caact aaaacactat ctgattctga tgaaacagaa cgagtttctg atgaaataaa 48ctaaa gaggctgtag aaatagcttg gaaagccaca gtaaaagtaa aagatgagtt 54atgta gaaaatgcag tcaaagaggc attggataaa ataaagacag aaaccgcgaa 6acaaaa cttacagata tagaagaagt agcagagtta gtattacaaa tagccaaaaa 66cggaa atagcgcaag aagttgtggc cttgttaaat actt 77Borrelia burgdorferi ctgctaa gtttattttt attgctctca atatctggtt ctttagataa tgaaggtgta 6aaaag attacgagtc aaaaaaacag agtatactag gtgaattaaa tcagctattg caaacta caaattcact aaaagaagca aaaaatacaa cagataattt aaatgcatca gaggcaa ataaagttgt agaagcagtt ataagtgtgg ttaatttaat ttcatctgct 24tcagg tgaaaggtca acaacaaata tgcacgattt agctcaaatg gcagaaatag 3agaaaa aataaaggaa tctagtgata aagtaatagt tgcggctaat gttgcgaaag 36tataa ccttactaaa gcagtagaac aaaatatgca aaaactgtac aaagagcaag 42caact aaaacactat ctgattctga tgaagcagaa cgagtttctg atgaaataaa 48ctaaa gaggctgtag aaatagcttg gaaagccaca gtaaaagtaa aagatgagtt 54atgta gaaaatgcag tcaaagaggc attggataaa ataaagacag aaaccgcgaa 6acaaaa cttacagata tagaagaagt agcagagtta gtattacaaa tagccaaaaa 66cggaa atagcgcaag aagttgtggc cttgttaaat actt 77Borrelia burgdorferi ctgttaa gtttattttt attgctctca atatcttgtt ctttagataa tgaaggtgta 6aaaag attacgagtc aaaaaaacag agtatactag gtgaattaaa tcagctattg caaacta caaattcact aaaagaagca aaaaatacaa cagataattt aaatgcatca gaggcaa ataaagttgt agaagcagtt ataagtgtgg ttaatttaat ttcatctgct 24tcagg tgaaaggtca acaacaaata tgcacgattt agctcaaatg gcagaaatag 3agaaaa aataaaggaa tctagtgata aagtaatagt tgcggctaat gttgcgaaag 36tataa ccttactaaa gcagtagaac aaaatatgca aaaactgtac aaagagcaag 42caact aaaacactat ctgattctga tgaagcagaa cgagtttctg atgaaataaa 48ctaaa gaggctgtag aaatagcttg gaaagccaca gtaaaagtaa aagatgagtt 54atgta gaaaatgcag tcaaagaggc attggataaa ataaagacag gaaccgcgaa 6acaaaa cttacagata tagaagaagt agcagagtta gtattacaaa tagccaaaaa 66cggaa atagcgcaag aagttgtggc cttgttaaat actt 7A Borrelia burgdorferi CDS (7 atg att atc aat cat aat aca tca gct att aat gct tca aga aat aat 48 Met Ile Ile Asn His Asn Thr Ser Ala Ile Asn Ala Ser Arg Asn Asn att aac gct gct aat ctt agt aaa act caa gaa aag ctt tct agt 96 Gly Ile Asn Ala Ala Asn Leu Ser Lys Thr Gln Glu Lys Leu Ser Ser 2 ggc tac aga att aat cga gct tct gat gat gct gct ggc atg gga gtt Tyr Arg Ile Asn Arg Ala Ser Asp Asp Ala Ala Gly Met Gly Val 35 4t ggt aag att aat gct caa ata aga ggt ttg tca caa gct tct aga Gly Lys Ile Asn Ala Gln Ile Arg Gly Leu Ser Gln Ala Ser Arg 5 aat act tca aag gct att aat ttt att cag aca aca gaa ggg aat tta 24hr Ser Lys Ala Ile Asn Phe Ile Gln Thr Thr Glu Gly Asn Leu 65 7 aat gaa gta gaa aaa gtc tta gta aga atg aag gaa ttg gca gtt caa 288 Asn Glu Val Glu Lys Val Leu Val Arg Met Lys Glu Leu Ala Val Gln 85 9a ggt aac ggc aca tat tca gat gca gac aga ggt tct ata caa att 336 Ser Gly Asn Gly Thr Tyr Ser Asp Ala Asp Arg Gly Ser Ile Gln Ile ata gag caa ctt aca gac gaa att aat aga att gct gat caa gct 384 Glu Ile Glu Gln Leu Thr Asp Glu Ile Asn Arg Ile Ala Asp Gln Ala tat aac caa atg cac atg tta tca aac aaa tct gct tct caa aat 432 Gln Tyr Asn Gln Met His Met Leu Ser Asn Lys Ser Ala Ser Gln Asn aga aca gct gaa gag ctt gga atg cag cct gca aaa att aac aca 48rg Thr Ala Glu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr cca gca tca ctt tca ggg ctt caa gcg tct tgg act tta aga gtt cat 528 Pro Ala Ser Leu Ser Gly Leu Gln Ala Ser Trp Thr Leu Arg Val His gga gca acc caa gat gaa gct att gct gta aat att tat gca gct 576 Val Gly Ala Thr Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ala Ala gtt gca aat ctt ttc tct ggt gag gga gct caa act gct cag gct 624 Asn Val Ala Asn Leu Phe Ser Gly Glu Gly Ala Gln Thr Ala Gln Ala 2ccg gtt caa gag ggt gtt caa cag gaa gga gct caa cag cca gca 672 Ala Pro Val Gln Glu Gly Val Gln Gln Glu Gly Ala Gln Gln Pro Ala 222ct aca gca cct tct caa ggc gga gtt aat tct cct gtt aat gtt 72la Thr Ala Pro Ser Gln Gly Gly Val Asn Ser Pro Val Asn Val 225 234ct aca gtt gat gct aat aca tca ctt gct aaa att gaa aat gct 768 Thr Thr Thr Val Asp Ala Asn Thr Ser Leu Ala Lys Ile Glu Asn Ala 245 25tt aga atg ata agt gat caa agg gca aat tta ggt gct ttc caa aat 8Arg Met Ile Ser Asp Gln Arg Ala Asn Leu Gly Ala Phe Gln Asn 267tt gaa tct ata aag aat agt act gag tat gca att gaa aat cta 864 Arg Leu Glu Ser Ile Lys Asn Ser Thr Glu Tyr Ala Ile Glu Asn Leu 275 28aa gca tct tat gct caa ata aaa gat gct aca atg aca gat gag gtt 9Ala Ser Tyr Ala Gln Ile Lys Asp Ala Thr Met Thr Asp Glu Val 29gca gca aca act aat atg att tta aca caa tct gca atg gca atg 96la Ala Thr Thr Asn Met Ile Leu Thr Gln Ser Ala Met Ala Met 33att gcg cag gct aat caa gtt ccc caa tat gtt ttg tca ttg ctt aga e Ala Gln Ala Asn Gln Val Pro Gln Tyr Val Leu Ser Leu Leu Arg 325 33aa PRT Borrelia burgdorferi Ile Ile Asn His Asn Thr Ser Ala Ile Asn Ala Ser Arg Asn Asn Ile Asn Ala Ala Asn Leu Ser Lys Thr Gln Glu Lys Leu Ser Ser 2 Gly Tyr Arg Ile Asn Arg Ala Ser Asp Asp Ala Ala Gly Met Gly Val 35 4r Gly Lys Ile Asn Ala Gln Ile Arg Gly Leu Ser Gln Ala Ser Arg 5 Asn Thr Ser Lys Ala Ile Asn Phe Ile Gln Thr Thr Glu Gly Asn Leu 65 7 Asn Glu Val Glu Lys Val Leu Val Arg Met Lys Glu Leu Ala Val Gln 85 9r Gly Asn Gly Thr Tyr Ser Asp Ala Asp Arg Gly Ser Ile Gln Ile Ile Glu Gln Leu Thr Asp Glu Ile Asn Arg Ile Ala Asp Gln Ala Tyr Asn Gln Met His Met Leu Ser Asn Lys Ser Ala Ser Gln Asn Arg Thr Ala Glu Glu Leu Gly Met Gln Pro Ala Lys Ile Asn Thr Pro Ala Ser Leu Ser Gly Leu Gln Ala Ser Trp Thr Leu Arg Val His Gly Ala Thr Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr Ala Ala Val Ala Asn Leu Phe Ser Gly Glu Gly Ala Gln Thr Ala Gln Ala 2Pro Val Gln Glu Gly Val Gln Gln Glu Gly Ala Gln Gln Pro Ala 222la Thr Ala Pro Ser Gln Gly Gly Val Asn Ser Pro Val Asn Val 225 234hr Thr Val Asp Ala Asn Thr Ser Leu Ala Lys Ile Glu Asn Ala 245 25le Arg Met Ile Ser Asp Gln Arg Ala Asn Leu Gly Ala Phe Gln Asn 267eu Glu Ser Ile Lys Asn Ser Thr Glu Tyr Ala Ile Glu Asn Leu 275 28ys Ala Ser Tyr Ala Gln Ile Lys Asp Ala Thr Met Thr Asp Glu Val 29Ala Ala Thr Thr Asn Met Ile Leu Thr Gln Ser Ala Met Ala Met 33Ile Ala Gln Ala Asn Gln Val Pro Gln Tyr Val Leu Ser Leu Leu Arg 325 3329 A Borrelia burgdorferi attatca atcataatac atcagctatt aatgcttcaa gaaataatgg cattaacgct 6tctta gtaaaactca agaaaagctt tctagtgggt acagaattaa tcgagcttct gatgctg ctggcatggg agtttctggt aagattaatg ctcaaataag aggtttgtca gcttcta gaaatacttc aaaggctatt aattttattc agacaacaga agggaattta 24agtag aaaaagtctt agtaagaatg aaggaattgg cagttcaatc aggtaacggc 3attcag atgcagacag aggttctata caaattgaaa tagagcaact tacagacgaa 36tagaa ttgctgatca agctcaatat aaccaaatgc acatgttatc aaacaaatct 42tcaaa atgtaagaac agctgaagag cttggaatgc agcctgcaaa aattaacaca 48atcac tttcagggtc tcaagcgtct tggactttaa gagttcatgt tggagcaaac 54tgaag ctattgctgt aaatatttat gcagctaatg ttgcaaatct tttctctggt 6gagctc aaactgctca ggctgcaccg gttcaagagg gtgttcaaca ggaaggagct 66gccag cacctgctac agcaccttct caaggcggag ttaattctcc tgttaatgtt 72tacag ttgatgctaa tacatcactt gctaaaattg aaaatgctat tagaatgata 78tcaaa gggcaaattt aggtgctttc caaaatagac ttgaatctat aaagaatagt 84gtatg caattgaaaa tctaaaagca tcttatgctc aaataaaaga tgctacaatg 9atgagg ttgtagcagc aacaactaat agtattttaa cacaatctgc aatggcaatg 96gcagg ctaatcaagt tccccaatat gttttgtcat tgcttaga DNA Borrelia burgdorferi attatca atcataatac atcagctatt aatgcttcaa gaaataatgc cattaatgct 6tctta gtaaaaccca agagaagctt tctagtggtt acagaattaa tcgagcttct gatgctg ctggtatggg ggtttctggc aagattaatg ctcaaataag aggcttatca gcttcta gaaacacttc aaaagctatc aattttattc agacaacaga aggaaattta 24agtag aaaaagtttt agtaagaatg aaagaattag cagttcaatc aggtaacgga 3attcag actcagacag aggttctata cagattgaaa tagagcaact tacagacgaa 36tagaa ttgctgatca ggctcaatat aaccaaatgc acatgttgtc aaacaaatct 42ccaaa atgtaaaaac agctgaagag cttggaatgc agcctgcaaa aattaacaca 48atcac tttcaggatc tcaagcttct tggactttaa gagttcatgt gggagcaaat 54tgaag caattgctgt aaatatttat tcagctaatg ttgcaaatct ttttgctggt 6gagctc aagctgctca ggctgcacct gttcaagagg gtgctcaaga agaaggagct 66accaa cacctgctac agcacctact caaggtggag ttaattctcc tgttaatgtt 72cacag ttgatgctaa tacatcactt gctaaaatag aaaatgctat tagaatgata 78tcaaa gagcaaattt aggtgctttc caaaatagac ttgaatctat aaagaatagc 84gtatg ctattgaaaa tctaaaagca tcttatgctc aaataaaaga tgctacaatg 9atgagg ttgtagcagc tacaactaat agtattttaa ctcaatctgc aatggcaatg 96acagg ctaatcaagt tcctcaatat gttttgtcat tgcttaga DNA Borrelia burgdorf eri 822 DNA Borrelia burgdorferi aaaaaat atttattggg aataggtcta atattagcct taatagcatg taagcaaaat 6cagcc ttgacgagaa aaacagcgtt tcagtagatt tgcctggtga aatgaaagtt gtaagca aagaaaaaaa caaagacggc aagtacgatc taattgcaac agtagacaag gagctta aaggaacttc tgataaaaac aatggatctg gagtacttga aggcgtaaaa 24caaaa gtaaagtaaa attaacaatt tctgacgatc taggtcaaac cacacttgaa 3tcaaag aagatggcaa aacactagta tcaaaaaaag taacttccaa agacaagtca 36agaag aaaaattcaa tgaaaaaggt gaagtatctg aaaaaataat aacaagagca 42aacca gacttgaata cacaggaatt aaaagcgatg gatctggaaa agctaaagag 48aaaag gctatgttct tgaaggaact ctaactgctg aaaaaacaac attggtggtt 54aggaa ctgttacttt aagcaaaaat

atttcaaaat ctggggaagt ttcagttgaa 6atgaca ctgacagtag tgctgctact aaaaaaactg cagcttggaa ttcaggcact 66tttaa caattactgt aaacagtaaa aaaactaaag accttgtgtt tacaaaagaa 72aatta cagtacaaca atacgactca aatggcacca aattagaggg gtcagcagtt 78tacaa aacttgatga aattaaaaac gctttaaaat aa 822 DNA Borrelia burgdorferi aaaaaat atttattggg aataggtcta atattagcct taatagcatg taagcaaaat 6cagcc ttgacgagaa aaacagcgtt tcagtagatt tgcctggtga aatgaacgtt gtaagca aagaaaaaaa caaagacggc aagtacgatc taattgcaac agtagacaag gagctta aaggaacttc tgataaaaac aatggatctg gagtacttga aggcgtaaaa 24caaaa gtaaagtaaa attaacaatt tctgacgatc taggtcaaac cacacttgaa 3tcaaag aagatggcaa aacactagta tcaaaaaaag taacttccaa agacaagtca 36agaag aaaaattcaa tgaaaaaggt gaagtatctg aaaaaataat aacaagagca 42aacca gacttgaata cacagaaatt aaaagcgatg gatctggaaa agctaaagag 48aaaag gctatgttct tgaaggaact ctaactgctg aaaaaacaac attggtggtt 54aggaa ctgttacttt aagcaaaaat atttcaaaat ctggggaagt ttcagttgaa 6atgaca ctgacagtag tgctgctact aaaaaaactg cagcttggaa ttcaggcact 66tttaa caattactgt aaacagtaaa aaaactaaag accttgtgtt tacaaaagaa 72aatta cagtacaaca atacgactca aatggcacca aattagaggg gtcagcagtt 78tacaa aacttgatga aattaaaaac gctttaaaat aa 822 DNA Borrelia burgdorferi aaaaaat atttattggg aataggtcta atattagcct taatagcatg taagcaaaat 6cagcc ttgacgagaa aaacagcgtt tcagtagatt tgcctggtga aatgaacgtt gtaagca aagaaaaaaa caaagacggc aagtacgatc taattgcaac agtagacaag gagctta aaggaacttc tgataaaaac aatggatctg gagtacttga aggcgtaaaa 24caaaa gtaaagtaaa attaacaatt tctgacgatc taggtcaaac cacacttgaa 3tcaaag aagatggcaa aacactagta tcaaaaaaag taacttccaa agacaagtca 36agaag aaaaattcaa tgaaaaaggt gaagtatctg aaaaaataat aacaagagca 42aacca gacttgaata cacagaaatt aaaagcgatg gatctggaaa agctaaagag 48aaaaa gctatgttct tgaaggaact ttaactgctg aaaaaacaac attggtggtt 54aggaa ctgttacttt aagcaaaaat atttcaaaat ctggggaagt ttcagttgaa 6atgaca ctgacagtag tgctgctact aaaaaaactg cagcttggaa ttcaggcact 66tttaa caattactgt aaacagtaaa aaaactaaag accttgtgtt tacaaaagaa 72aatta cagtacaaca atacgactca aatggcacca aattagaggg gtcagcagtt 78tacaa aacttgatga aattaaaaac gctttaaaat aa 822 DNA Borrelia burgdorferi aaaaaat atttattggg aataggtcta atattagcct taatagcatg taagcaaaat 6cagcc ttgatgagaa aaacagcgtt tcagtagatt tacctggtga aatgaaagtt gtaagca aagaaaaaga caaagatggt aaatacagtc taatggcaac agtagacaag gagctta aaggaacttc tgataaaagc aacggttctg gaacacttga aggtgaaaaa 24caaaa gtaaagcaaa attaacaatt tctgaagatc taagtaaaac cacatttgaa 3tcaaag aagatggcaa aacattagta tcaaaaaaag taaattctaa agataagtca 36agaag aaaaattcaa cgcaaaaggt gaattatctg aaaaaacaat actaagagca 42aacca ggcttgaata cacagaaata aaaagcgatg gaaccggaaa agctaaagaa 48aaaag actttgctct tgaaggaact ctagctgccg acaaaacaac attgaaagtt 54aggca ctgttgtttt aagcaaacac attccaaact ctggagaaat aacagttgag 6atgact ctaactctac tcaggctact aaaaaaactg gaaaatggga ttcaaatact 66tttaa caattagtgt gaatagcaaa aaaactaaaa acattgtatt tacaaaagaa 72aataa cagtacaaaa atacgactca gcaggcacca atctagaagg caacgcagtc 78taaaa cacttgatga acttaaaaac gctttaaaat a 822orrelia burgdorferi aaaaaat atttattggg aataggtcta atattagcct taatagcatg taagcaaaat 6cagcc ttgatgaaaa aaatagcgtt tcagtagatt tacctggtgg aatgaaagtt gtaagta aagaaaaaga caaagatggt aaatacagtc taatggcaac agtagaaaag gagctta aaggaacttc tgataaaaac aacggttctg gaacacttga aggtgaaaaa 24caaaa gtaaagtaaa attaacaatt gctgaggatc taagtaaaac cacatttgaa 3tcaaag aagatggcaa aacattagta tcgaaaaaag taacccttaa agacaagtca 36agaag aaaaattcaa cgaaaagggt gaaatatctg aaaaaacaat agtaagagca 42aacca gacttgaata cacagacata aaaagcgata aaaccggaaa agctaaagaa 48aaaag actttactct tgaaggaact ctagctgctg acggcaaaac aacattgaaa 54agagg gcactgttac tttaagcaag aacatttcaa aatccggaga aataacagtt 6ttgatg acactgactc tagcggcaat aaaaaatccg gaacatggga ttcaggtact 66tttaa caattagtaa aaacagacaa aaaactaaac aacttgtatt cacaaaagaa 72aataa cagtacaaaa ctacgactca gcaggcacca atctagaagg caaagcagtc 78tacaa cacttaaaga acttaaaaac gctttaaaat a 8225 DNA Artificial Sequence CDS (25) Chimeric Nucleic Acid aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gat gaa aaa aat agc gtt tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 gat tta cct ggt gga atg aca gtt ctt gta agt aaa gaa aaa gac aaa Leu Pro Gly Gly Met Thr Val Leu Val Ser Lys Glu Lys Asp Lys 35 4c ggt aaa tac agt cta gag gca aca gta gac aag ctt gag ctt aaa Gly Lys Tyr Ser Leu Glu Ala Thr Val Asp Lys Leu Glu Leu Lys 5 gga act tct gat aaa aac aac ggt tct gga aca ctt gaa ggt gaa aaa 24hr Ser Asp Lys Asn Asn Gly Ser Gly Thr Leu Glu Gly Glu Lys 65 7 act gac aaa agt aaa gta aaa tta aca att gct gat gac cta agt caa 288 Thr Asp Lys Ser Lys Val Lys Leu Thr Ile Ala Asp Asp Leu Ser Gln 85 9t aaa ttt gaa att ttc aaa gaa gat gcc aaa aca tta gta tca aaa 336 Thr Lys Phe Glu Ile Phe Lys Glu Asp Ala Lys Thr Leu Val Ser Lys gta acc ctt aaa gac aag tca tca aca gaa gaa aaa ttc aac gaa 384 Lys Val Thr Leu Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu ggt gaa aca tct gaa aaa aca ata gta aga gca aat gga acc aga 432 Lys Gly Glu Thr Ser Glu Lys Thr Ile Val Arg Ala Asn Gly Thr Arg gaa tac aca gac ata aaa agc gat gga tcc gga aaa gct aaa gaa 48lu Tyr Thr Asp Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu gtt tta aaa gac ttt act ctt gaa gga act cta gct gct gac ggc aaa 528 Val Leu Lys Asp Phe Thr Leu Glu Gly Thr Leu Ala Ala Asp Gly Lys aca ttg aaa gtt aca gaa ggc act gtt gtt tta agc aag aac att 576 Thr Thr Leu Lys Val Thr Glu Gly Thr Val Val Leu Ser Lys Asn Ile aaa tcc gga gaa ata aca gtt gca ctt gat gac tct gac act act 624 Leu Lys Ser Gly Glu Ile Thr Val Ala Leu Asp Asp Ser Asp Thr Thr 2gct act aaa aaa act gga aaa tgg gat tca aat act tcc act tta 672 Gln Ala Thr Lys Lys Thr Gly Lys Trp Asp Ser Asn Thr Ser Thr Leu 222tt agt gtg aat agc aaa aaa act aaa aac att gta ttt aca aaa 72le Ser Val Asn Ser Lys Lys Thr Lys Asn Ile Val Phe Thr Lys 225 234ac aca ata aca gta caa aaa tac gac tca gca ggc acc aat cta 768 Glu Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu 245 25aa ggc aac gca gtc gaa att aaa aca ctt gat gaa ctt aaa aac gct 8Gly Asn Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala 267aa taa 825 Leu Lys * PRT Artificial Sequence Chimeric Protein Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 Asp Leu Pro Gly Gly Met Thr Val Leu Val Ser Lys Glu Lys Asp Lys 35 4p Gly Lys Tyr Ser Leu Glu Ala Thr Val Asp Lys Leu Glu Leu Lys 5 Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Thr Leu Glu Gly Glu Lys 65 7 Thr Asp Lys Ser Lys Val Lys Leu Thr Ile Ala Asp Asp Leu Ser Gln 85 9r Lys Phe Glu Ile Phe Lys Glu Asp Ala Lys Thr Leu Val Ser Lys Val Thr Leu Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Gly Glu Thr Ser Glu Lys Thr Ile Val Arg Ala Asn Gly Thr Arg Glu Tyr Thr Asp Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys Asp Phe Thr Leu Glu Gly Thr Leu Ala Ala Asp Gly Lys Thr Leu Lys Val Thr Glu Gly Thr Val Val Leu Ser Lys Asn Ile Lys Ser Gly Glu Ile Thr Val Ala Leu Asp Asp Ser Asp Thr Thr 2Ala Thr Lys Lys Thr Gly Lys Trp Asp Ser Asn Thr Ser Thr Leu 222le Ser Val Asn Ser Lys Lys Thr Lys Asn Ile Val Phe Thr Lys 225 234sp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu 245 25lu Gly Asn Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala 267ys DNA Artificial Sequence CDS (22) Chimeric Nucleic Acid aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gat gaa aaa aac agc gct tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Ala Ser Val 2 gat ttg cct ggt gag atg aaa gtt ctt gta agt aaa gaa aaa gac aaa Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asp Lys 35 4c ggt aag tac agt cta aag gca aca gta gac aag att gag cta aaa Gly Lys Tyr Ser Leu Lys Ala Thr Val Asp Lys Ile Glu Leu Lys 5 gga act tct gat aaa gac aat ggt tct gga gtg ctt gaa ggt aca aaa 24hr Ser Asp Lys Asp Asn Gly Ser Gly Val Leu Glu Gly Thr Lys 65 7 gat gac aaa agt aaa gca aaa tta aca att gct gac gat cta agt aaa 288 Asp Asp Lys Ser Lys Ala Lys Leu Thr Ile Ala Asp Asp Leu Ser Lys 85 9c aca ttc gaa ctt tta aaa gaa gat ggc aaa aca tta gtg tca aga 336 Thr Thr Phe Glu Leu Leu Lys Glu Asp Gly Lys Thr Leu Val Ser Arg gta agt tct aga gac aaa aca tca aca gat gaa atg ttc aat gaa 384 Lys Val Ser Ser Arg Asp Lys Thr Ser Thr Asp Glu Met Phe Asn Glu ggt gaa ttg tct gca aaa acc atg aca aga gaa aat gga acc aaa 432 Lys Gly Glu Leu Ser Ala Lys Thr Met Thr Arg Glu Asn Gly Thr Lys gaa tat aca gaa atg aaa agc gat gga acc gga aaa gct aaa gaa 48lu Tyr Thr Glu Met Lys Ser Asp Gly Thr Gly Lys Ala Lys Glu gtt tta aaa aag ttt act ctt gaa gga aaa gta gct aat gat aaa gta 528 Val Leu Lys Lys Phe Thr Leu Glu Gly Lys Val Ala Asn Asp Lys Val ttg gaa gta aaa gaa gga acc gtt act tta agt aag gaa att gca 576 Thr Leu Glu Val Lys Glu Gly Thr Val Thr Leu Ser Lys Glu Ile Ala tct gga gaa gta aca gtt gct ctt aat gac act aac act act cag 624 Lys Ser Gly Glu Val Thr Val Ala Leu Asn Asp Thr Asn Thr Thr Gln 2act aaa aaa act ggc gca tgg gat tca aaa act tct act tta aca 672 Ala Thr Lys Lys Thr Gly Ala Trp Asp Ser Lys Thr Ser Thr Leu Thr 222gt gtt aac agc aaa aaa act aca caa ctt gtg ttt act aaa caa 72er Val Asn Ser Lys Lys Thr Thr Gln Leu Val Phe Thr Lys Gln 225 234ca ata act gta caa aaa tac gac tcc gca ggt acc aat tta gaa 768 Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25gc aca gca gtc gaa att aaa aca ctt gat gaa ctt aaa aac gct tta 8Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu 267aa 822 Lys * PRT Artificial Sequence Chimeric Protein Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Ala Ser Val 2 Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asp Lys 35 4p Gly Lys Tyr Ser Leu Lys Ala Thr Val Asp Lys Ile Glu Leu Lys 5 Gly Thr Ser Asp Lys Asp Asn Gly Ser Gly Val Leu Glu Gly Thr Lys 65 7 Asp Asp Lys Ser Lys Ala Lys Leu Thr Ile Ala Asp Asp Leu Ser Lys 85 9r Thr Phe Glu Leu Leu Lys Glu Asp Gly Lys Thr Leu Val Ser Arg Val Ser Ser Arg Asp Lys Thr Ser Thr Asp Glu Met Phe Asn Glu Gly Glu Leu Ser Ala Lys Thr Met Thr Arg Glu Asn Gly Thr Lys Glu Tyr Thr Glu Met Lys Ser Asp Gly Thr Gly Lys Ala Lys Glu Val Leu Lys Lys Phe Thr Leu Glu Gly Lys Val Ala Asn Asp Lys Val Leu Glu Val Lys Glu Gly Thr Val Thr Leu Ser Lys Glu Ile Ala Ser Gly Glu Val Thr Val Ala Leu Asn Asp Thr Asn Thr Thr Gln 2Thr Lys Lys Thr Gly Ala Trp Asp Ser Lys Thr Ser Thr Leu Thr 222er Val Asn Ser Lys Lys Thr Thr Gln Leu Val Phe Thr Lys Gln 225 234hr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25ly Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu 2674NA Artificial Sequence CDS (22) Chimeric Nucleic Acid aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gat gaa aaa aac agc gct tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Ala Ser Val 2 gat ttg cct ggt gag atg aaa gtt ctt gta agt aaa gaa aaa gac aaa Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asp Lys 35 4c ggt aag tac agt cta aag gca aca gta gac aag att gag cta aaa Gly Lys Tyr Ser Leu Lys Ala Thr Val Asp Lys Ile Glu Leu Lys 5 gga act tct gat aaa gac aat ggt tct gga gtg ctt gaa ggt aca aaa 24hr Ser Asp Lys Asp Asn Gly Ser Gly Val Leu Glu Gly Thr Lys 65 7 gat gac aaa agt aaa gca aaa tta aca att gct gac gat cta agt aaa 288 Asp Asp Lys Ser Lys Ala Lys Leu Thr Ile Ala Asp Asp Leu Ser Lys 85 9c aca ttc gaa ctt tta aaa gaa gat ggc aaa aca tta gtg tca aga 336 Thr Thr Phe Glu Leu Leu Lys Glu Asp Gly Lys Thr Leu Val Ser Arg gta agt tct aga gac aaa aca tca aca gat gaa atg ttc aat gaa 384 Lys Val Ser Ser Arg Asp Lys Thr Ser Thr Asp Glu Met Phe Asn Glu ggt gaa ttg tct gca aaa acc atg aca aga gaa aat gga acc aaa 432 Lys Gly Glu Leu Ser Ala Lys Thr Met Thr Arg Glu Asn Gly Thr Lys gaa tat aca gaa atg aaa agc gat gga acc gga aaa gct aaa gaa 48lu Tyr Thr Glu Met Lys Ser Asp Gly Thr Gly Lys Ala Lys Glu gtt tta aaa aag ttt act ctt gaa gga aaa gta gct aat gat aaa gta 528 Val Leu Lys Lys Phe Thr Leu Glu Gly Lys Val Ala Asn Asp Lys Val ttg gaa gta aaa gaa gga acc gtt act tta agt aag gaa att tca 576 Thr Leu Glu Val Lys Glu Gly Thr Val Thr Leu Ser Lys Glu Ile Ser tct ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt gct 624 Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2act aaa aaa act gca gct tgg aat tca aaa act tcc act tta aca 672 Ala Thr Lys Lys Thr Ala Ala

Trp Asn Ser Lys Thr Ser Thr Leu Thr 222gt gtg aat agc caa aaa acc aaa aac ctt gta ttc aca aaa gaa 72er Val Asn Ser Gln Lys Thr Lys Asn Leu Val Phe Thr Lys Glu 225 234ca ata aca gta caa aaa tac gac tca gca ggc acc aat cta gaa 768 Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25gc aaa gca gtc gaa att aca aca ctt aaa gaa ctt aaa aac gct tta 8Lys Ala Val Glu Ile Thr Thr Leu Lys Glu Leu Lys Asn Ala Leu 267aa 822 Lys * PRT Artificial Sequence Chimeric Protein Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Ala Ser Val 2 Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asp Lys 35 4p Gly Lys Tyr Ser Leu Lys Ala Thr Val Asp Lys Ile Glu Leu Lys 5 Gly Thr Ser Asp Lys Asp Asn Gly Ser Gly Val Leu Glu Gly Thr Lys 65 7 Asp Asp Lys Ser Lys Ala Lys Leu Thr Ile Ala Asp Asp Leu Ser Lys 85 9r Thr Phe Glu Leu Leu Lys Glu Asp Gly Lys Thr Leu Val Ser Arg Val Ser Ser Arg Asp Lys Thr Ser Thr Asp Glu Met Phe Asn Glu Gly Glu Leu Ser Ala Lys Thr Met Thr Arg Glu Asn Gly Thr Lys Glu Tyr Thr Glu Met Lys Ser Asp Gly Thr Gly Lys Ala Lys Glu Val Leu Lys Lys Phe Thr Leu Glu Gly Lys Val Ala Asn Asp Lys Val Leu Glu Val Lys Glu Gly Thr Val Thr Leu Ser Lys Glu Ile Ser Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr 222er Val Asn Ser Gln Lys Thr Lys Asn Leu Val Phe Thr Lys Glu 225 234hr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25ly Lys Ala Val Glu Ile Thr Thr Leu Lys Glu Leu Lys Asn Ala Leu 26743 822 DNA Artificial Sequence CDS (22) Chimeric Nucleic Acid aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gac gag aaa aac agc gtt tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 gat ttg cct ggt gaa atg aaa gtt ctt gta agc aaa gaa aaa aac aaa Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys 35 4c ggc aag tac gat cta att gca aca gta gac aag ctt gag ctt aaa Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys 5 gga act tct gat aaa aac aat gga tct gga gta ctt gaa ggc gta aaa 24hr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 gct gac aaa agt aaa gta aaa tta aca att tct gac gat cta ggt caa 288 Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln 85 9c aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca cta gta tca aaa 336 Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys gta act tcc aaa gac aag tca tca aca gaa gaa aaa ttc aat gaa 384 Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu ggt gaa gta tct gaa aaa ata ata aca aga gca aat gga acc aaa 432 Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asn Gly Thr Lys gaa tat aca gaa atg aaa agc gat gga acc gga aaa gct aaa gaa 48lu Tyr Thr Glu Met Lys Ser Asp Gly Thr Gly Lys Ala Lys Glu gtt tta aaa aag ttt act ctt gaa gga aaa gta gct aat gat aaa gta 528 Val Leu Lys Lys Phe Thr Leu Glu Gly Lys Val Ala Asn Asp Lys Val ttg gaa gta aaa gaa gga acc gtt act tta agt aag gaa att tca 576 Thr Leu Glu Val Lys Glu Gly Thr Val Thr Leu Ser Lys Glu Ile Ser tct ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt gct 624 Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2act aaa aaa act gca gct tgg aat tca aaa act tcc act tta aca 672 Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr 222gt gtg aat agc caa aaa acc aaa aac ctt gta ttc aca aaa gaa 72er Val Asn Ser Gln Lys Thr Lys Asn Leu Val Phe Thr Lys Glu 225 234ca ata aca gta caa aaa tac gac tca gca ggc acc aat cta gaa 768 Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25gc aaa gca gtc gaa att aca aca ctt aaa gaa ctt aaa aac gct tta 8Lys Ala Val Glu Ile Thr Thr Leu Lys Glu Leu Lys Asn Ala Leu 267aa 822 Lys * PRT Artificial Sequence Chimeric Protein Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys 35 4p Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys 5 Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln 85 9r Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asn Gly Thr Lys Glu Tyr Thr Glu Met Lys Ser Asp Gly Thr Gly Lys Ala Lys Glu Val Leu Lys Lys Phe Thr Leu Glu Gly Lys Val Ala Asn Asp Lys Val Leu Glu Val Lys Glu Gly Thr Val Thr Leu Ser Lys Glu Ile Ser Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr 222er Val Asn Ser Gln Lys Thr Lys Asn Leu Val Phe Thr Lys Glu 225 234hr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25ly Lys Ala Val Glu Ile Thr Thr Leu Lys Glu Leu Lys Asn Ala Leu 26745 A Artificial Sequence CDS (362) Chimeric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata aat aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gaa att gct gct aaa gct att ggt aaa aaa Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 ata cac caa aat aat ggt ttg gat acc gaa aat aat cac aat gga tca 24is Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 ttg tta gcg gga gct tat gca ata tca acc cta ata aaa caa aaa tta 288 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9t gga ttg aaa aat gaa gga tta aag gaa aaa att gat gcg gct aag 336 Asp Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys tgt tct gaa aca ttt act aat aaa tta aaa gaa aaa cac aca gat 384 Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp ggt aaa gaa ggt gtt act gat gct gat gca aaa gaa gcc att tta 432 Leu Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu gca aat ggt act aaa act aaa ggt gct gaa gaa ctt gga aaa tta 48la Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu ttt gaa tca gta gag gtc ttg tca aaa gca gct aaa gag atg ctt gct 528 Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala tca gtt aaa gag ctt aca agc cct gtt gtg gca gaa agt cca aaa 576 Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys cct tcc atg gcc aag caa aat gtt agc agc ctt gac gag aaa aac 624 Lys Pro Ser Met Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn 2gtt tca gta gat ttg cct ggt gaa atg aaa gtt ctt gta agc aaa 672 Ser Val Ser Val Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys 222aa aac aaa gac ggc aag tac gat cta att gca aca gta gac aag 72ys Asn Lys Asp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys 225 234ag ctt aaa gga act tct gat aaa aac aat gga tct gga gta ctt 768 Leu Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu 245 25aa ggc gta aaa gct gac aaa agt aaa gta aaa tta aca att tct gac 8Gly Val Lys Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp 267ta ggt caa acc aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca 864 Asp Leu Gly Gln Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr 275 28ta gta tca aaa aaa gta act tcc aaa gac aag tca tca aca gaa gaa 9Val Ser Lys Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu 29ttc aat gaa aaa ggt gaa gta tct gaa aaa ata ata aca aga gca 96he Asn Glu Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala 33gac gga acc aga ctt gaa tac aca gga att aaa agc gat gga tct gga p Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly 325 33aa gct aaa gag gtt tta aaa ggc tat gtt ctt gaa gga act cta act s Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr 345aa aaa aca aca ttg gtg gtt aaa gaa gga act gtt act tta agc a Glu Lys Thr Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser 355 36aa aat att tca aaa tct ggg gaa gtt tca gtt gaa ctt aat gac act s Asn Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr 378gt agt gct gct act aaa aaa act gca gct tgg aat tca ggc act p Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Gly Thr 385 39act tta aca att act gta aac agt aaa aaa act aaa gac ctt gtg r Thr Leu Thr Ile Thr Val Asn Ser Lys Lys Thr Lys Asp Leu Val 44aca aaa gaa aac aca att aca gta caa caa tac gac tca aat ggc e Thr Lys Glu Asn Thr Ile Thr Val Gln Gln Tyr Asp Ser Asn Gly 423aa tta gag ggg tca gca gtt gaa att aca aaa ctt gat gaa att r Lys Leu Glu Gly Ser Ala Val Glu Ile Thr Lys Leu Asp Glu Ile 435 44aa aac gct tta aaa taa s Asn Ala Leu Lys * 4553 PRT Artificial Sequence Chimeric Protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 Ile His Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9p Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu Ala Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys Pro Ser Met Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn 2Val Ser Val Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys 222ys Asn Lys Asp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys 225 234lu Leu Lys Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu 245 25lu Gly Val Lys Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp 267eu Gly Gln Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr 275 28eu Val Ser Lys Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu 29Phe Asn Glu Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala 33Asp Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly 325 33ys Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr 345lu Lys Thr Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser 355 36ys Asn Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr 378er Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Gly Thr 385 39Thr Leu Thr Ile Thr Val Asn Ser Lys Lys Thr Lys Asp Leu Val 44Thr Lys Glu Asn Thr Ile Thr Val Gln Gln Tyr Asp Ser Asn Gly 423ys Leu Glu Gly Ser Ala Val Glu Ile Thr Lys Leu Asp Glu Ile 435 44ys Asn Ala Leu Lys 45353 DNA Artificial Sequence CDS (353) Chimeric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata aat aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gaa att gct gct aaa gct att ggt aaa aaa Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 ata cac caa aat aat ggt ttg gat acc gaa aat aat cac aat gga tca 24is Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn

His Asn Gly Ser 65 7 ttg tta gcg gga gct tat gca ata tca acc cta ata aaa caa aaa tta 288 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9t gga ttg aaa aat gaa gga tta aag gaa aaa att gat gcg gct aag 336 Asp Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys tgt tct gaa aca ttt act aat aaa tta aaa gaa aaa cac aca gat 384 Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp ggt aaa gaa ggt gtt act gat gct gat gca aaa gaa gcc att tta 432 Leu Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu gca aat ggt act aaa act aaa ggt gct gaa gaa ctt gga aaa tta 48la Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu ttt gaa tca gta gag gtc ttg tca aaa gca gct aaa gag atg ctt gct 528 Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala tca gtt aaa gag ctt aca agc cct gtt gtg gca gaa agt cca gcc 576 Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Ala gcc aag caa aat gtt agc agc ctt gac gag aaa aac agc gtt tca 624 Met Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser 2gat ttg cct ggt gaa atg aaa gtt ctt gta agc aaa gaa aaa aac 672 Val Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn 222ac ggc aag tac gat cta att gca aca gta gac aag ctt gag ctt 72sp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu 225 234ga act tct gat aaa aac aat gga tct gga gta ctt gaa ggc gta 768 Lys Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val 245 25aa gct gac aaa agt aaa gta aaa tta aca att tct gac gat cta ggt 8Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly 267cc aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca cta gta tca 864 Gln Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser 275 28aa aaa gta act tcc aaa gac aag tca tca aca gaa gaa aaa ttc aat 9Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn 29aaa ggt gaa gta tct gaa aaa ata ata aca aga gca gac gga acc 96ys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr 33aga ctt gaa tac aca gga att aaa agc gat gga tct gga aaa gct aaa g Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys 325 33ag gtt tta aaa ggc tat gtt ctt gaa gga act cta act gct gaa aaa u Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys 345ca ttg gtg gtt aaa gaa gga act gtt act tta agc aaa aat att r Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile 355 36ca aaa tct ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt r Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser 378ct act aaa aaa act gca gct tgg aat tca ggc act tca act tta a Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Gly Thr Ser Thr Leu 385 39att act gta aac agt aaa aaa act aaa gac ctt gtg ttt aca aaa r Ile Thr Val Asn Ser Lys Lys Thr Lys Asp Leu Val Phe Thr Lys 44aac aca att aca gta caa caa tac gac tca aat ggc acc aaa tta u Asn Thr Ile Thr Val Gln Gln Tyr Asp Ser Asn Gly Thr Lys Leu 423gg tca gca gtt gaa att aca aaa ctt gat gaa att aaa aac gct u Gly Ser Ala Val Glu Ile Thr Lys Leu Asp Glu Ile Lys Asn Ala 435 44ta aaa taa u Lys * 455rtificial Sequence Chimeric Protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 Ile His Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9p Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu Ala Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Ala Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser 2Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn 222sp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu 225 234ly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val 245 25ys Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly 267hr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser 275 28ys Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn 29Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr 33Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys 325 33lu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys 345hr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile 355 36er Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser 378la Thr Lys Lys Thr Ala Ala Trp Asn Ser Gly Thr Ser Thr Leu 385 39Ile Thr Val Asn Ser Lys Lys Thr Lys Asp Leu Val Phe Thr Lys 44Asn Thr Ile Thr Val Gln Gln Tyr Asp Ser Asn Gly Thr Lys Leu 423ly Ser Ala Val Glu Ile Thr Lys Leu Asp Glu Ile Lys Asn Ala 435 44eu Lys 4534rtificial Sequence CDS (34eric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata agt aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gag ctt gct aaa gct att ggt aaa aaa ata Leu Ser Ser Ile Asp Glu Leu Ala Lys Ala Ile Gly Lys Lys Ile 5 aaa aac gat ggt agt tta gat aat gaa gca aat cgc aac gag tca ttg 24sn Asp Gly Ser Leu Asp Asn Glu Ala Asn Arg Asn Glu Ser Leu 65 7 tta gca gga gct tat aca ata tca acc tta ata aca caa aaa tta agt 288 Leu Ala Gly Ala Tyr Thr Ile Ser Thr Leu Ile Thr Gln Lys Leu Ser 85 9a tta aac gga tca gaa ggt tta aag gaa aag att gcc gca gct aag 336 Lys Leu Asn Gly Ser Glu Gly Leu Lys Glu Lys Ile Ala Ala Ala Lys tgc tct gaa gag ttt agt act aaa cta aaa gat aat cat gca cag 384 Lys Cys Ser Glu Glu Phe Ser Thr Lys Leu Lys Asp Asn His Ala Gln ggt ata cag ggc gtt act gat gaa aat gca aaa aaa gct att tta 432 Leu Gly Ile Gln Gly Val Thr Asp Glu Asn Ala Lys Lys Ala Ile Leu gca aat gca gcg ggt aaa gat aag ggc gtt gaa gaa ctt gaa aag 48la Asn Ala Ala Gly Lys Asp Lys Gly Val Glu Glu Leu Glu Lys ttg tcc gga tca tta gaa agc tta tca aaa gca gct aaa gag atg ctt 528 Leu Ser Gly Ser Leu Glu Ser Leu Ser Lys Ala Ala Lys Glu Met Leu aat tca gtt aaa gag ctt aca agc cct gtt gtc cat ggc aag caa 576 Ala Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val His Gly Lys Gln gtt agc agc ctt gac gag aaa aac agc gtt tca gta gat ttg cct 624 Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val Asp Leu Pro 2gaa atg aaa gtt ctt gta agc aaa gaa aaa aac aaa gac ggc aag 672 Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys Asp Gly Lys 222at cta att gca aca gta gac aag ctt gag ctt aaa gga act tct 72sp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys Gly Thr Ser 225 234aa aac aat gga tct gga gta ctt gaa ggc gta aaa gct gac aaa 768 Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys Ala Asp Lys 245 25gt aaa gta aaa tta aca att tct gac gat cta ggt caa acc aca ctt 8Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln Thr Thr Leu 267tt ttc aaa gaa gat ggc aaa aca cta gta tca aaa aaa gta act 864 Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Lys Val Thr 275 28cc aaa gac aag tca tca aca gaa gaa aaa ttc aat gaa aaa ggt gaa 9Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Lys Gly Glu 29tct gaa aaa ata ata aca aga gca gac gga acc aga ctt gaa tac 96er Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg Leu Glu Tyr 33aca gga att aaa agc gat gga tct gga aaa gct aaa gag gtt tta aaa r Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys 325 33gc tat gtt ctt gaa gga act cta act gct gaa aaa aca aca ttg gtg y Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr Thr Leu Val 345aa gaa gga act gtt act tta agc aaa aat att tca aaa tct ggg l Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser Lys Ser Gly 355 36aa gtt tca gtt gaa ctt aat gac act gac agt agt gct gct act aaa u Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala Ala Thr Lys 378ct gca gct tgg aat tca ggc act tca act tta aca att act gta s Thr Ala Ala Trp Asn Ser Gly Thr Ser Thr Leu Thr Ile Thr Val 385 39agt aaa aaa act aaa gac ctt gtg ttt aca aaa gaa aac aca att n Ser Lys Lys Thr Lys Asp Leu Val Phe Thr Lys Glu Asn Thr Ile 44gta caa caa tac gac tca aat ggc acc aaa tta gag ggg tca gca r Val Gln Gln Tyr Asp Ser Asn Gly Thr Lys Leu Glu Gly Ser Ala 423aa att aca aaa ctt gat gaa att aaa aac gct tta aaa taa l Glu Ile Thr Lys Leu Asp Glu Ile Lys Asn Ala Leu Lys * 435 445RT Artificial Sequence Chimeric Protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Leu Ala Lys Ala Ile Gly Lys Lys Ile 5 Lys Asn Asp Gly Ser Leu Asp Asn Glu Ala Asn Arg Asn Glu Ser Leu 65 7 Leu Ala Gly Ala Tyr Thr Ile Ser Thr Leu Ile Thr Gln Lys Leu Ser 85 9s Leu Asn Gly Ser Glu Gly Leu Lys Glu Lys Ile Ala Ala Ala Lys Cys Ser Glu Glu Phe Ser Thr Lys Leu Lys Asp Asn His Ala Gln Gly Ile Gln Gly Val Thr Asp Glu Asn Ala Lys Lys Ala Ile Leu Ala Asn Ala Ala Gly Lys Asp Lys Gly Val Glu Glu Leu Glu Lys Leu Ser Gly Ser Leu Glu Ser Leu Ser Lys Ala Ala Lys Glu Met Leu Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val His Gly Lys Gln Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val Asp Leu Pro 2Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys Asp Gly Lys 222sp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys Gly Thr Ser 225 234ys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys Ala Asp Lys 245 25er Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln Thr Thr Leu 267al Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Lys Val Thr 275 28er Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Lys Gly Glu 29Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg Leu Glu Tyr 33Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys 325 33ly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr Thr Leu Val 345ys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser Lys Ser Gly 355 36lu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala Ala Thr Lys 378hr Ala Ala Trp Asn Ser Gly Thr Ser Thr Leu Thr Ile Thr Val 385 39Ser Lys Lys Thr Lys Asp Leu Val Phe Thr Lys Glu Asn Thr Ile 44Val Gln Gln Tyr Asp Ser Asn Gly Thr Lys Leu Glu Gly Ser Ala 423lu Ile Thr Lys Leu Asp Glu Ile Lys Asn Ala Leu Lys 435 445DNA Artificial Sequence CDS (362) Chimeric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata aat aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gaa att gct gct aaa gct att ggt aaa aaa Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 ata cac caa aat aat ggt ttg gat acc gaa aat aat cac aat gga tca 24is Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 ttg tta gcg gga gct tat gca ata tca acc cta ata aaa caa aaa tta 288 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9t gga ttg aaa aat gaa gga tta aag gaa aaa att gat gcg gct aag 336 Asp Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys tgt tct

gaa aca ttt act aat aaa tta aaa gaa aaa cac aca gat 384 Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp ggt aaa gaa ggt gtt act gat gct gat gca aaa gaa gcc att tta 432 Leu Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu gca aat ggt act aaa act aaa ggt gct gaa gaa ctt gga aaa tta 48la Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu ttt gaa tca gta gag gtc ttg tca aaa gca gct aaa gag atg ctt gct 528 Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala tca gtt aaa gag ctt aca agc cct gtt gtg gca gaa agt cca aaa 576 Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys cct tcc atg gcc aag caa aat gtt agc agc ctt gac gag aaa aac 624 Lys Pro Ser Met Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn 2gtt tca gta gat ttg cct ggt gaa atg aaa gtt ctt gta agc aaa 672 Ser Val Ser Val Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys 222aa aac aaa gac ggc aag tac gat cta att gca aca gta gac aag 72ys Asn Lys Asp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys 225 234ag ctt aaa gga act tct gat aaa aac aat gga tct gga gta ctt 768 Leu Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu 245 25aa ggc gta aaa gct gac aaa agt aaa gta aaa tta aca att tct gac 8Gly Val Lys Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp 267ta ggt caa acc aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca 864 Asp Leu Gly Gln Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr 275 28ta gta tca aaa aaa gta act tcc aaa gac aag tca tca aca gaa gaa 9Val Ser Lys Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu 29ttc aat gaa aaa ggt gaa gta tct gaa aaa ata ata aca aga gca 96he Asn Glu Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala 33gac gga acc aga ctt gaa tac aca gga att aaa agc gat gga tct gga p Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly 325 33aa gct aaa gag gtt tta aaa ggc tat gtt ctt gaa gga act cta act s Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr 345aa aaa aca aca ttg gtg gtt aaa gaa gga act gtt act tta agc a Glu Lys Thr Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser 355 36aa aat att tca aaa tct ggg gaa gtt tca gtt gaa ctt aat gac act s Asn Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr 378gt agt gct gct act aaa aaa act gca gct tgg aat tca aaa act p Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr 385 39act tta aca att agt gtg aat agc caa aaa acc aaa aac ctt gta r Thr Leu Thr Ile Ser Val Asn Ser Gln Lys Thr Lys Asn Leu Val 44aca aaa gaa gac aca ata aca gta caa aaa tac gac tca gca ggc e Thr Lys Glu Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly 423at cta gaa ggc aaa gca gtc gaa att aca aca ctt aaa gaa ctt r Asn Leu Glu Gly Lys Ala Val Glu Ile Thr Thr Leu Lys Glu Leu 435 44aa aac gct tta aaa taa s Asn Ala Leu Lys * 4553 PRT Artificial Sequence Chimeric Protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 Ile His Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9p Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu Ala Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys Pro Ser Met Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn 2Val Ser Val Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys 222ys Asn Lys Asp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys 225 234lu Leu Lys Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu 245 25lu Gly Val Lys Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp 267eu Gly Gln Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr 275 28eu Val Ser Lys Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu 29Phe Asn Glu Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala 33Asp Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly 325 33ys Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr 345lu Lys Thr Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser 355 36ys Asn Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr 378er Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr 385 39Thr Leu Thr Ile Ser Val Asn Ser Gln Lys Thr Lys Asn Leu Val 44Thr Lys Glu Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly 423sn Leu Glu Gly Lys Ala Val Glu Ile Thr Thr Leu Lys Glu Leu 435 44ys Asn Ala Leu Lys 4534rtificial Sequence CDS (34eric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata agt aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gag ctt gct aaa gct att ggt aaa aaa ata Leu Ser Ser Ile Asp Glu Leu Ala Lys Ala Ile Gly Lys Lys Ile 5 aaa aac gat ggt agt tta gat aat gaa gca aat cgc aac gag tca ttg 24sn Asp Gly Ser Leu Asp Asn Glu Ala Asn Arg Asn Glu Ser Leu 65 7 tta gca gga gct tat aca ata tca acc tta ata aca caa aaa tta agt 288 Leu Ala Gly Ala Tyr Thr Ile Ser Thr Leu Ile Thr Gln Lys Leu Ser 85 9a tta aac gga tca gaa ggt tta aag gaa aag att gcc gca gct aag 336 Lys Leu Asn Gly Ser Glu Gly Leu Lys Glu Lys Ile Ala Ala Ala Lys tgc tct gaa gag ttt agt act aaa cta aaa gat aat cat gca cag 384 Lys Cys Ser Glu Glu Phe Ser Thr Lys Leu Lys Asp Asn His Ala Gln ggt ata cag ggc gtt act gat gaa aat gca aaa aaa gct att tta 432 Leu Gly Ile Gln Gly Val Thr Asp Glu Asn Ala Lys Lys Ala Ile Leu gca aat gca gcg ggt aaa gat aag ggc gtt gaa gaa ctt gaa aag 48la Asn Ala Ala Gly Lys Asp Lys Gly Val Glu Glu Leu Glu Lys ttg tcc gga tca tta gaa agc tta tca aaa gca gct aaa gag atg ctt 528 Leu Ser Gly Ser Leu Glu Ser Leu Ser Lys Ala Ala Lys Glu Met Leu aat tca gtt aaa gag ctt aca agc cct gtt gtc cat ggc aag caa 576 Ala Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val His Gly Lys Gln gtt agc agc ctt gac gag aaa aac agc gtt tca gta gat ttg cct 624 Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val Asp Leu Pro 2gaa atg aaa gtt ctt gta agc aaa gaa aaa aac aaa gac ggc aag 672 Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys Asp Gly Lys 222at cta att gca aca gta gac aag ctt gag ctt aaa gga act tct 72sp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys Gly Thr Ser 225 234aa aac aat gga tct gga gta ctt gaa ggc gta aaa gct gac aaa 768 Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys Ala Asp Lys 245 25gt aaa gta aaa tta aca att tct gac gat cta ggt caa acc aca ctt 8Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln Thr Thr Leu 267tt ttc aaa gaa gat ggc aaa aca cta gta tca aaa aaa gta act 864 Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Lys Val Thr 275 28cc aaa gac aag tca tca aca gaa gaa aaa ttc aat gaa aaa ggt gaa 9Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Lys Gly Glu 29tct gaa aaa ata ata aca aga gca gac gga acc aga ctt gaa tac 96er Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg Leu Glu Tyr 33aca gga att aaa agc gat gga tct gga aaa gct aaa gag gtt tta aaa r Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys 325 33gc tat gtt ctt gaa gga act cta act gct gaa aaa aca aca ttg gtg y Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr Thr Leu Val 345aa gaa gga act gtt act tta agc aaa aat att tca aaa tct ggg l Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser Lys Ser Gly 355 36aa gtt tca gtt gaa ctt aat gac act gac agt agt gct gct act aaa u Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala Ala Thr Lys 378ct gca gct tgg aat tca aaa act tcc act tta aca att agt gtg s Thr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr Ile Ser Val 385 39agc caa aaa acc aaa aac ctt gta ttc aca aaa gaa gac aca ata n Ser Gln Lys Thr Lys Asn Leu Val Phe Thr Lys Glu Asp Thr Ile 44gta caa aaa tac gac tca gca ggc acc aat cta gaa ggc aaa gca r Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu Gly Lys Ala 423aa att aca aca ctt aaa gaa ctt aaa aac gct tta aaa taa l Glu Ile Thr Thr Leu Lys Glu Leu Lys Asn Ala Leu Lys * 435 4454 446 PRT Artificial Sequence Chimeric protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Leu Ala Lys Ala Ile Gly Lys Lys Ile 5 Lys Asn Asp Gly Ser Leu Asp Asn Glu Ala Asn Arg Asn Glu Ser Leu 65 7 Leu Ala Gly Ala Tyr Thr Ile Ser Thr Leu Ile Thr Gln Lys Leu Ser 85 9s Leu Asn Gly Ser Glu Gly Leu Lys Glu Lys Ile Ala Ala Ala Lys Cys Ser Glu Glu Phe Ser Thr Lys Leu Lys Asp Asn His Ala Gln Gly Ile Gln Gly Val Thr Asp Glu Asn Ala Lys Lys Ala Ile Leu Ala Asn Ala Ala Gly Lys Asp Lys Gly Val Glu Glu Leu Glu Lys Leu Ser Gly Ser Leu Glu Ser Leu Ser Lys Ala Ala Lys Glu Met Leu Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val His Gly Lys Gln Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val Asp Leu Pro 2Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys Asp Gly Lys 222sp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys Gly Thr Ser 225 234ys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys Ala Asp Lys 245 25er Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln Thr Thr Leu 267al Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Lys Val Thr 275 28er Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Lys Gly Glu 29Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg Leu Glu Tyr 33Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys 325 33ly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr Thr Leu Val 345ys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser Lys Ser Gly 355 36lu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala Ala Thr Lys 378hr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr Ile Ser Val 385 39Ser Gln Lys Thr Lys Asn Leu Val Phe Thr Lys Glu Asp Thr Ile 44Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu Gly Lys Ala 423lu Ile Thr Thr Leu Lys Glu Leu Lys Asn Ala Leu Lys 435 4455 A Artificial Sequence CDS (362) Chimeric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata aat aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gaa att gct gct aaa gct att ggt aaa aaa Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 ata cac caa aat aat ggt ttg gat acc gaa aat aat cac aat gga tca 24is Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 ttg tta gcg gga gct tat gca ata tca acc cta ata aaa caa aaa tta 288 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9t gga ttg aaa aat gaa gga tta aag gaa aaa att gat gcg gct aag 336 Asp Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys tgt tct gaa aca ttt act aat aaa tta aaa gaa aaa cac aca gat 384 Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp ggt aaa gaa ggt gtt act gat gct gat gca aaa gaa gcc att tta 432 Leu Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu gca aat ggt act aaa act aaa ggt gct gaa gaa ctt gga aaa tta 48la Asn Gly

Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu ttt gaa tca gta gag gtc ttg tca aaa gca gct aaa gag atg ctt gct 528 Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala tca gtt aaa gag ctt aca agc cct gtt gtg gca gaa agt cca aaa 576 Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys cct tcc atg gcc aag caa aat gtt agc agc ctt gac gag aaa aac 624 Lys Pro Ser Met Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn 2gtt tca gta gat ttg cct ggt gaa atg aaa gtt ctt gta agc aaa 672 Ser Val Ser Val Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys 222aa aac aaa gac ggc aag tac gat cta att gca aca gta gac aag 72ys Asn Lys Asp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys 225 234ag ctt aaa gga act tct gat aaa aac aat gga tct gga gta ctt 768 Leu Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu 245 25aa ggc gta aaa gct gac aaa agt aaa gta aaa tta aca att tct gac 8Gly Val Lys Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp 267ta ggt caa acc aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca 864 Asp Leu Gly Gln Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr 275 28ta gta tca aaa aaa gta act tcc aaa gac aag tca tca aca gaa gaa 9Val Ser Lys Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu 29ttc aat gaa aaa ggt gaa gta tct gaa aaa ata ata aca aga gca 96he Asn Glu Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala 33gac gga acc aga ctt gaa tac aca gga att aaa agc gat gga tct gga p Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly 325 33aa gct aaa gag gtt tta aaa ggc tat gtt ctt gaa gga act cta act s Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr 345aa aaa aca aca ttg gtg gtt aaa gaa gga act gtt act tta agc a Glu Lys Thr Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser 355 36aa aat att tca aaa tct ggg gaa gtt tca gtt gaa ctt aat gac act s Asn Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr 378gt agt gct gct act aaa aaa act gca gct tgg aat tca aaa act p Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr 385 39act tta aca att agt gtt aac agc aaa aaa act aca caa ctt gtg r Thr Leu Thr Ile Ser Val Asn Ser Lys Lys Thr Thr Gln Leu Val 44act aaa caa gac aca ata act gta caa aaa tac gac tcc gca ggt e Thr Lys Gln Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly 423at tta gaa ggc aca gca gtc gaa att aaa aca ctt gat gaa ctt r Asn Leu Glu Gly Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu 435 44aa aac gct tta aaa taa s Asn Ala Leu Lys * 4553 PRT Artificial Sequence Chimeric Protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 Ile His Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9p Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu Ala Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys Pro Ser Met Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn 2Val Ser Val Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys 222ys Asn Lys Asp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys 225 234lu Leu Lys Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu 245 25lu Gly Val Lys Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp 267eu Gly Gln Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr 275 28eu Val Ser Lys Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu 29Phe Asn Glu Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala 33Asp Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly 325 33ys Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr 345lu Lys Thr Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser 355 36ys Asn Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr 378er Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr 385 39Thr Leu Thr Ile Ser Val Asn Ser Lys Lys Thr Thr Gln Leu Val 44Thr Lys Gln Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly 423sn Leu Glu Gly Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu 435 44ys Asn Ala Leu Lys 4534rtificial Sequence CDS (34eric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata agt aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gag ctt gct aaa gct att ggt aaa aaa ata Leu Ser Ser Ile Asp Glu Leu Ala Lys Ala Ile Gly Lys Lys Ile 5 aaa aac gat ggt agt tta gat aat gaa gca aat cgc aac gag tca ttg 24sn Asp Gly Ser Leu Asp Asn Glu Ala Asn Arg Asn Glu Ser Leu 65 7 tta gca gga gct tat aca ata tca acc tta ata aca caa aaa tta agt 288 Leu Ala Gly Ala Tyr Thr Ile Ser Thr Leu Ile Thr Gln Lys Leu Ser 85 9a tta aac gga tca gaa ggt tta aag gaa aag att gcc gca gct aag 336 Lys Leu Asn Gly Ser Glu Gly Leu Lys Glu Lys Ile Ala Ala Ala Lys tgc tct gaa gag ttt agt act aaa cta aaa gat aat cat gca cag 384 Lys Cys Ser Glu Glu Phe Ser Thr Lys Leu Lys Asp Asn His Ala Gln ggt ata cag ggc gtt act gat gaa aat gca aaa aaa gct att tta 432 Leu Gly Ile Gln Gly Val Thr Asp Glu Asn Ala Lys Lys Ala Ile Leu gca aat gca gcg ggt aaa gat aag ggc gtt gaa gaa ctt gaa aag 48la Asn Ala Ala Gly Lys Asp Lys Gly Val Glu Glu Leu Glu Lys ttg tcc gga tca tta gaa agc tta tca aaa gca gct aaa gag atg ctt 528 Leu Ser Gly Ser Leu Glu Ser Leu Ser Lys Ala Ala Lys Glu Met Leu aat tca gtt aaa gag ctt aca agc cct gtt gtc cat ggc aag caa 576 Ala Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val His Gly Lys Gln gtt agc agc ctt gac gag aaa aac agc gtt tca gta gat ttg cct 624 Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val Asp Leu Pro 2gaa atg aaa gtt ctt gta agc aaa gaa aaa aac aaa gac ggc aag 672 Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys Asp Gly Lys 222at cta att gca aca gta gac aag ctt gag ctt aaa gga act tct 72sp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys Gly Thr Ser 225 234aa aac aat gga tct gga gta ctt gaa ggc gta aaa gct gac aaa 768 Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys Ala Asp Lys 245 25gt aaa gta aaa tta aca att tct gac gat cta ggt caa acc aca ctt 8Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln Thr Thr Leu 267tt ttc aaa gaa gat ggc aaa aca cta gta tca aaa aaa gta act 864 Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Lys Val Thr 275 28cc aaa gac aag tca tca aca gaa gaa aaa ttc aat gaa aaa ggt gaa 9Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Lys Gly Glu 29tct gaa aaa ata ata aca aga gca gac gga acc aga ctt gaa tac 96er Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg Leu Glu Tyr 33aca gga att aaa agc gat gga tct gga aaa gct aaa gag gtt tta aaa r Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys 325 33gc tat gtt ctt gaa gga act cta act gct gaa aaa aca aca ttg gtg y Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr Thr Leu Val 345aa gaa gga act gtt act tta agc aaa aat att tca aaa tct ggg l Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser Lys Ser Gly 355 36aa gtt tca gtt gaa ctt aat gac act gac agt agt gct gct act aaa u Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala Ala Thr Lys 378ct gca gct tgg aat tca aaa act tct act tta aca att agt gtt s Thr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr Ile Ser Val 385 39agc aaa aaa act aca caa ctt gtg ttt act aaa caa tac aca ata n Ser Lys Lys Thr Thr Gln Leu Val Phe Thr Lys Gln Tyr Thr Ile 44gta aaa caa tac gac tcc gca ggt acc aat tta gaa ggc aca gca r Val Lys Gln Tyr Asp Ser Ala Gly Thr Asn Leu Glu Gly Thr Ala 423aa att aaa aca ctt gat gaa ctt aaa aac gct tta aaa taa l Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu Lys * 435 4458 446 PRT Artificial Sequence Chimeric protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Leu Ala Lys Ala Ile Gly Lys Lys Ile 5 Lys Asn Asp Gly Ser Leu Asp Asn Glu Ala Asn Arg Asn Glu Ser Leu 65 7 Leu Ala Gly Ala Tyr Thr Ile Ser Thr Leu Ile Thr Gln Lys Leu Ser 85 9s Leu Asn Gly Ser Glu Gly Leu Lys Glu Lys Ile Ala Ala Ala Lys Cys Ser Glu Glu Phe Ser Thr Lys Leu Lys Asp Asn His Ala Gln Gly Ile Gln Gly Val Thr Asp Glu Asn Ala Lys Lys Ala Ile Leu Ala Asn Ala Ala Gly Lys Asp Lys Gly Val Glu Glu Leu Glu Lys Leu Ser Gly Ser Leu Glu Ser Leu Ser Lys Ala Ala Lys Glu Met Leu Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val His Gly Lys Gln Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val Asp Leu Pro 2Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys Asp Gly Lys 222sp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys Gly Thr Ser 225 234ys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys Ala Asp Lys 245 25er Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln Thr Thr Leu 267al Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Lys Val Thr 275 28er Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Lys Gly Glu 29Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg Leu Glu Tyr 33Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys 325 33ly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr Thr Leu Val 345ys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser Lys Ser Gly 355 36lu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala Ala Thr Lys 378hr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr Ile Ser Val 385 39Ser Lys Lys Thr Thr Gln Leu Val Phe Thr Lys Gln Tyr Thr Ile 44Val Lys Gln Tyr Asp Ser Ala Gly Thr Asn Leu Glu Gly Thr Ala 423lu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu Lys 435 4459 A Artificial Sequence CDS (365) Chimeric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata agt aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gaa att gct gct aaa gct att ggt aaa aaa Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 ata cac caa aat aat ggt ttg gat acc gaa tat aat cac aat gga tca 24is Gln Asn Asn Gly Leu Asp Thr Glu Tyr Asn His Asn Gly Ser 65 7 ttg tta gcg gga gct tat gca ata tca acc cta ata aaa caa aaa tta 288 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9t gga ttg aaa aat gaa gga tta aag gaa aaa att gat gcg gct aag 336 Asp Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys tgt tct gaa aca ttt act aat aaa tta aaa gaa aaa cac aca gat 384 Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp ggt aaa gaa ggt gtt act gat gct gat gca aaa gaa gcc att tta 432 Leu Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu aca aat ggt act aaa act aaa ggt gct gaa gaa ctt gga aaa tta 48hr Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu ttt gaa tca gta gag gtc ttg tca aaa gca gct aaa gag atg ctt gct 528 Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala tca gtt aaa gag ctt aca agc cct gtt gtg gca gaa agt cca aaa 576 Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys

cct tcc atg gcc aag caa aat gtt agc agc ctt gat gaa aaa aat 624 Lys Pro Ser Met Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn 2gtt tca gta gat tta cct ggt gga atg aca gtt ctt gta agt aaa 672 Ser Val Ser Val Asp Leu Pro Gly Gly Met Thr Val Leu Val Ser Lys 222aa gac aaa gac ggt aaa tac agt cta gag gca aca gta gac aag 72ys Asp Lys Asp Gly Lys Tyr Ser Leu Glu Ala Thr Val Asp Lys 225 234ag ctt aaa gga act tct gat aaa aac aac ggt tct gga aca ctt 768 Leu Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Thr Leu 245 25aa ggt gaa aaa act gac aaa agt aaa gta aaa tta aca att gct gat 8Gly Glu Lys Thr Asp Lys Ser Lys Val Lys Leu Thr Ile Ala Asp 267ta agt caa act aaa ttt gaa att ttc aaa gaa gat gcc aaa aca 864 Asp Leu Ser Gln Thr Lys Phe Glu Ile Phe Lys Glu Asp Ala Lys Thr 275 28ta gta tca aaa aaa gta acc ctt aaa gac aag tca tca aca gaa gaa 9Val Ser Lys Lys Val Thr Leu Lys Asp Lys Ser Ser Thr Glu Glu 29ttc aac gaa aag ggt gaa aca tct gaa aaa aca ata gta aga gca 96he Asn Glu Lys Gly Glu Thr Ser Glu Lys Thr Ile Val Arg Ala 33aat gga acc aga ctt gaa tac aca gac ata aaa agc gat gga tcc gga n Gly Thr Arg Leu Glu Tyr Thr Asp Ile Lys Ser Asp Gly Ser Gly 325 33aa gct aaa gaa gtt tta aaa gac ttt act ctt gaa gga act cta gct s Ala Lys Glu Val Leu Lys Asp Phe Thr Leu Glu Gly Thr Leu Ala 345ac ggc aaa aca aca ttg aaa gtt aca gaa ggc act gtt gtt tta a Asp Gly Lys Thr Thr Leu Lys Val Thr Glu Gly Thr Val Val Leu 355 36gc aag aac att tta aaa tcc gga gaa ata aca gtt gca ctt gat gac r Lys Asn Ile Leu Lys Ser Gly Glu Ile Thr Val Ala Leu Asp Asp 378ac act act cag gct act aaa aaa act gga aaa tgg gat tca aat r Asp Thr Thr Gln Ala Thr Lys Lys Thr Gly Lys Trp Asp Ser Asn 385 39tcc act tta aca att agt gtg aat agc aaa aaa act aaa aac att r Ser Thr Leu Thr Ile Ser Val Asn Ser Lys Lys Thr Lys Asn Ile 44ttt aca aaa gaa gac aca ata aca gta caa aaa tac gac tca gca l Phe Thr Lys Glu Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala 423cc aat cta gaa ggc aac gca gtc gaa att aaa aca ctt gat gaa y Thr Asn Leu Glu Gly Asn Ala Val Glu Ile Lys Thr Leu Asp Glu 435 44tt aaa aac gct tta aaa tag u Lys Asn Ala Leu Lys * 4554 PRT Artificial Sequence Chimeric protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 Ile His Gln Asn Asn Gly Leu Asp Thr Glu Tyr Asn His Asn Gly Ser 65 7 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9p Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu Thr Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys Pro Ser Met Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn 2Val Ser Val Asp Leu Pro Gly Gly Met Thr Val Leu Val Ser Lys 222ys Asp Lys Asp Gly Lys Tyr Ser Leu Glu Ala Thr Val Asp Lys 225 234lu Leu Lys Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Thr Leu 245 25lu Gly Glu Lys Thr Asp Lys Ser Lys Val Lys Leu Thr Ile Ala Asp 267eu Ser Gln Thr Lys Phe Glu Ile Phe Lys Glu Asp Ala Lys Thr 275 28eu Val Ser Lys Lys Val Thr Leu Lys Asp Lys Ser Ser Thr Glu Glu 29Phe Asn Glu Lys Gly Glu Thr Ser Glu Lys Thr Ile Val Arg Ala 33Asn Gly Thr Arg Leu Glu Tyr Thr Asp Ile Lys Ser Asp Gly Ser Gly 325 33ys Ala Lys Glu Val Leu Lys Asp Phe Thr Leu Glu Gly Thr Leu Ala 345sp Gly Lys Thr Thr Leu Lys Val Thr Glu Gly Thr Val Val Leu 355 36er Lys Asn Ile Leu Lys Ser Gly Glu Ile Thr Val Ala Leu Asp Asp 378sp Thr Thr Gln Ala Thr Lys Lys Thr Gly Lys Trp Asp Ser Asn 385 39Ser Thr Leu Thr Ile Ser Val Asn Ser Lys Lys Thr Lys Asn Ile 44Phe Thr Lys Glu Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala 423hr Asn Leu Glu Gly Asn Ala Val Glu Ile Lys Thr Leu Asp Glu 435 44eu Lys Asn Ala Leu Lys 45344 DNA Artificial Sequence CDS (344) Chimeric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata agt aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gag ctt gct aaa gct att ggt aaa aaa ata Leu Ser Ser Ile Asp Glu Leu Ala Lys Ala Ile Gly Lys Lys Ile 5 aaa aac gat ggt agt tta gat aat gaa gca aat cgc aac gag tca ttg 24sn Asp Gly Ser Leu Asp Asn Glu Ala Asn Arg Asn Glu Ser Leu 65 7 tta gca gga gct tat aca ata tca acc tta ata aca caa aaa tta agt 288 Leu Ala Gly Ala Tyr Thr Ile Ser Thr Leu Ile Thr Gln Lys Leu Ser 85 9a tta aac gga tca gaa ggt tta aag gaa aag att gcc gca gct aag 336 Lys Leu Asn Gly Ser Glu Gly Leu Lys Glu Lys Ile Ala Ala Ala Lys tgc tct gaa gag ttt agt act aaa cta aaa gat aat cat gca cag 384 Lys Cys Ser Glu Glu Phe Ser Thr Lys Leu Lys Asp Asn His Ala Gln ggt ata cag ggc gtt act gat gaa aat gca aaa aaa gct att tta 432 Leu Gly Ile Gln Gly Val Thr Asp Glu Asn Ala Lys Lys Ala Ile Leu gca aat gca gcg ggt aaa gat aag ggc gtt gaa gaa ctt gaa aag 48la Asn Ala Ala Gly Lys Asp Lys Gly Val Glu Glu Leu Glu Lys ttg tcc gga tca tta gaa agc tta tca aaa gca gct aaa gag atg ctt 528 Leu Ser Gly Ser Leu Glu Ser Leu Ser Lys Ala Ala Lys Glu Met Leu aat tca gtt aaa gag ctt aca agc cct gtt gtc cat ggc aag caa 576 Ala Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val His Gly Lys Gln gtt agc agc ctt gat gaa aaa aat agc gtt tca gta gat tta cct 624 Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val Asp Leu Pro 2gga atg aca gtt ctt gta agt aaa gaa aaa gac aaa gac ggt aaa 672 Gly Gly Met Thr Val Leu Val Ser Lys Glu Lys Asp Lys Asp Gly Lys 222gt cta gag gca aca gta gac aag ctt gag ctt aaa gga act tct 72er Leu Glu Ala Thr Val Asp Lys Leu Glu Leu Lys Gly Thr Ser 225 234aa aac aac ggt tct gga aca ctt gaa ggt gaa aaa act gac aaa 768 Asp Lys Asn Asn Gly Ser Gly Thr Leu Glu Gly Glu Lys Thr Asp Lys 245 25gt aaa gta aaa tta aca att gct gat gac cta agt caa act aaa ttt 8Lys Val Lys Leu Thr Ile Ala Asp Asp Leu Ser Gln Thr Lys Phe 267tt ttc aaa gaa gat gcc aaa aca tta gta tca aaa aaa gta acc 864 Glu Ile Phe Lys Glu Asp Ala Lys Thr Leu Val Ser Lys Lys Val Thr 275 28tt aaa gac aag tca tca aca gaa gaa aaa ttc aac gaa aag ggt gaa 9Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Lys Gly Glu 29tct gaa aaa aca ata gta aga gca aat gga acc aga ctt gaa tac 96er Glu Lys Thr Ile Val Arg Ala Asn Gly Thr Arg Leu Glu Tyr 33aca gac ata aaa agc gat gga tcc gga aaa gct aaa gaa gtt tta aaa r Asp Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys 325 33ac ttt act ctt gaa gga act cta gct gct gac ggc aaa aca aca ttg p Phe Thr Leu Glu Gly Thr Leu Ala Ala Asp Gly Lys Thr Thr Leu 345tt aca gaa ggc act gtt gtt tta agc aag aac att tta aaa tcc s Val Thr Glu Gly Thr Val Val Leu Ser Lys Asn Ile Leu Lys Ser 355 36ga gaa ata aca gtt gca ctt gat gac tct gac act act cag gct act y Glu Ile Thr Val Ala Leu Asp Asp Ser Asp Thr Thr Gln Ala Thr 378aa act gga aaa tgg gat tca aat act tcc act tta aca att agt s Lys Thr Gly Lys Trp Asp Ser Asn Thr Ser Thr Leu Thr Ile Ser 385 39aat agc aaa aaa act aaa aac att gta ttt aca aaa gaa gac aca l Asn Ser Lys Lys Thr Lys Asn Ile Val Phe Thr Lys Glu Asp Thr 44aca gta caa aaa tac gac tca gca ggc acc aat cta gaa ggc aac e Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu Gly Asn 423tc gaa att aaa aca ctt gat gaa ctt aaa aac gct tta aaa tag a Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu Lys * 435 4462 447 PRT Artificial Sequence Chimeric protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Leu Ala Lys Ala Ile Gly Lys Lys Ile 5 Lys Asn Asp Gly Ser Leu Asp Asn Glu Ala Asn Arg Asn Glu Ser Leu 65 7 Leu Ala Gly Ala Tyr Thr Ile Ser Thr Leu Ile Thr Gln Lys Leu Ser 85 9s Leu Asn Gly Ser Glu Gly Leu Lys Glu Lys Ile Ala Ala Ala Lys Cys Ser Glu Glu Phe Ser Thr Lys Leu Lys Asp Asn His Ala Gln Gly Ile Gln Gly Val Thr Asp Glu Asn Ala Lys Lys Ala Ile Leu Ala Asn Ala Ala Gly Lys Asp Lys Gly Val Glu Glu Leu Glu Lys Leu Ser Gly Ser Leu Glu Ser Leu Ser Lys Ala Ala Lys Glu Met Leu Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val His Gly Lys Gln Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val Asp Leu Pro 2Gly Met Thr Val Leu Val Ser Lys Glu Lys Asp Lys Asp Gly Lys 222er Leu Glu Ala Thr Val Asp Lys Leu Glu Leu Lys Gly Thr Ser 225 234ys Asn Asn Gly Ser Gly Thr Leu Glu Gly Glu Lys Thr Asp Lys 245 25er Lys Val Lys Leu Thr Ile Ala Asp Asp Leu Ser Gln Thr Lys Phe 267le Phe Lys Glu Asp Ala Lys Thr Leu Val Ser Lys Lys Val Thr 275 28eu Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Lys Gly Glu 29Ser Glu Lys Thr Ile Val Arg Ala Asn Gly Thr Arg Leu Glu Tyr 33Thr Asp Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys 325 33sp Phe Thr Leu Glu Gly Thr Leu Ala Ala Asp Gly Lys Thr Thr Leu 345al Thr Glu Gly Thr Val Val Leu Ser Lys Asn Ile Leu Lys Ser 355 36ly Glu Ile Thr Val Ala Leu Asp Asp Ser Asp Thr Thr Gln Ala Thr 378ys Thr Gly Lys Trp Asp Ser Asn Thr Ser Thr Leu Thr Ile Ser 385 39Asn Ser Lys Lys Thr Lys Asn Ile Val Phe Thr Lys Glu Asp Thr 44Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu Gly Asn 423al Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu Lys 435 4463 A Artificial Sequence CDS (3meric nucleic acid gct tgt aat aat tca gga aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata agt aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 att aca gaa tct aac gca gtt gtt ctg gct gtg aaa gaa att gaa act Thr Glu Ser Asn Ala Val Val Leu Ala Val Lys Glu Ile Glu Thr 35 4g ctt gca tct ata gat gaa ctt gct act aaa gct att ggt aaa aaa Leu Ala Ser Ile Asp Glu Leu Ala Thr Lys Ala Ile Gly Lys Lys 5 ata caa caa aat ggt ggt tta gct gtc gaa gcg ggg cat aat gga aca 24ln Gln Asn Gly Gly Leu Ala Val Glu Ala Gly His Asn Gly Thr 65 7 ttg tta gca ggt gct tat aca ata tca aaa cta ata aca caa aaa tta 288 Leu Leu Ala Gly Ala Tyr Thr Ile Ser Lys Leu Ile Thr Gln Lys Leu 85 9t gga ttg aaa aat tca gaa aaa tta aag gaa aaa att gaa aat gct 336 Asp Gly Leu Lys Asn Ser Glu Lys Leu Lys Glu Lys Ile Glu Asn Ala aaa tgt tct gaa gat ttt act aaa aaa cta gaa gga gaa cat gcg 384 Lys Lys Cys Ser Glu Asp Phe Thr Lys Lys Leu Glu Gly Glu His Ala ctt gga att gaa aat gtt act gat gag aat gca aaa aaa gct att 432 Gln Leu Gly Ile Glu Asn Val Thr Asp Glu Asn Ala Lys Lys Ala Ile ata aca gat gca gct aaa gat aag ggc gct gca gag ctt gaa aag 48le Thr Asp Ala Ala Lys Asp Lys Gly Ala Ala Glu Leu Glu Lys cta ttt aaa gca gta gaa aac ttg gca aaa gca gct aaa gag atg ctt 528 Leu Phe Lys Ala Val Glu Asn Leu Ala Lys Ala Ala Lys Glu Met Leu aat tca gtt aaa gag ctt aca agt cct att gtg cat ggc gtt tca 576 Ala Asn Ser Val Lys Glu Leu Thr Ser Pro Ile Val His Gly Val Ser gat ttg cct ggt gaa atg aaa gtt ctt gta agc aaa gaa aaa aac 624 Val Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn 2gac ggc aag tac gat cta att gca aca gta gac aag ctt gag ctt 672 Lys Asp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu 222ga act tct gat aaa aac aat gga

tct gga gta ctt gaa ggc gta 72ly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val 225 234ct gac aaa agt aaa gta aaa tta aca att tct gac gat cta ggt 768 Lys Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly 245 25aa acc aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca cta gta tca 8Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser 267aa gta act tcc aaa gac aag tca tca aca gaa gaa aaa ttc aat 864 Lys Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn 275 28aa aaa ggt gaa gta tct gaa aaa ata ata aca aga gca gac gga acc 9Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr 29ctt gaa tac aca gga att aaa agc gat gga tct gga aaa gct aaa 96eu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys 33gag gtt tta aaa aaa ttt act ctt gaa gga aaa gta gct aat gat aaa u Val Leu Lys Lys Phe Thr Leu Glu Gly Lys Val Ala Asn Asp Lys 325 33ta aca ttg gaa gta aaa gaa gga acc gtt act tta agt aag aac att l Thr Leu Glu Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile 345aa tct ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt r Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser 355 36ct gct act aaa aaa act gca gct tgg aat tca ggc act tca act tta a Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Gly Thr Ser Thr Leu 378tt act gta aac agt aaa aaa act aaa gac ctt gtg ttt aca aaa r Ile Thr Val Asn Ser Lys Lys Thr Lys Asp Leu Val Phe Thr Lys 385 39aac aca att aca gta caa caa tac gac tca aat ggc acc aaa tta u Asn Thr Ile Thr Val Gln Gln Tyr Asp Ser Asn Gly Thr Lys Leu 44ggg tca gca gtt gaa att aca aaa ctt gat gaa att aaa aac gct u Gly Ser Ala Val Glu Ile Thr Lys Leu Asp Glu Ile Lys Asn Ala 423aa taa u Lys * PRT Artificial Sequence Chimeric protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 Ile Thr Glu Ser Asn Ala Val Val Leu Ala Val Lys Glu Ile Glu Thr 35 4u Leu Ala Ser Ile Asp Glu Leu Ala Thr Lys Ala Ile Gly Lys Lys 5 Ile Gln Gln Asn Gly Gly Leu Ala Val Glu Ala Gly His Asn Gly Thr 65 7 Leu Leu Ala Gly Ala Tyr Thr Ile Ser Lys Leu Ile Thr Gln Lys Leu 85 9p Gly Leu Lys Asn Ser Glu Lys Leu Lys Glu Lys Ile Glu Asn Ala Lys Cys Ser Glu Asp Phe Thr Lys Lys Leu Glu Gly Glu His Ala Leu Gly Ile Glu Asn Val Thr Asp Glu Asn Ala Lys Lys Ala Ile Ile Thr Asp Ala Ala Lys Asp Lys Gly Ala Ala Glu Leu Glu Lys Leu Phe Lys Ala Val Glu Asn Leu Ala Lys Ala Ala Lys Glu Met Leu Asn Ser Val Lys Glu Leu Thr Ser Pro Ile Val His Gly Val Ser Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn 2Asp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu 222ly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val 225 234la Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly 245 25ln Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser 267ys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn 275 28lu Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr 29Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys 33Glu Val Leu Lys Lys Phe Thr Leu Glu Gly Lys Val Ala Asn Asp Lys 325 33al Thr Leu Glu Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile 345ys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser 355 36la Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Gly Thr Ser Thr Leu 378le Thr Val Asn Ser Lys Lys Thr Lys Asp Leu Val Phe Thr Lys 385 39Asn Thr Ile Thr Val Gln Gln Tyr Asp Ser Asn Gly Thr Lys Leu 44Gly Ser Ala Val Glu Ile Thr Lys Leu Asp Glu Ile Lys Asn Ala 423ys 2 DNA Artificial Sequence CDS (332) Chimeric nucleic acid gct tgt agt aat tca ggg aaa ggt ggg gat tct gca tct act aat 48 Met Ala Cys Ser Asn Ser Gly Lys Gly Gly Asp Ser Ala Ser Thr Asn gct gac gag tct gcg aaa ggg cct aat ctt aca gaa ata agc aaa 96 Pro Ala Asp Glu Ser Ala Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys 2 aaa att aca gat tct aat gca ttt gta ctt gct gtt aaa gaa gtt gag Ile Thr Asp Ser Asn Ala Phe Val Leu Ala Val Lys Glu Val Glu 35 4t ttg gtt tta tct ata gat gaa ctt gct aag aaa gct att ggt caa Leu Val Leu Ser Ile Asp Glu Leu Ala Lys Lys Ala Ile Gly Gln 5 aaa ata gac aat aat aat ggt tta gct gct tta aat aat cag aat gga 24le Asp Asn Asn Asn Gly Leu Ala Ala Leu Asn Asn Gln Asn Gly 65 7 tcg ttg tta gca gga gcc tat gca ata tca acc cta ata aca gaa aaa 288 Ser Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Thr Glu Lys 85 9g agt aaa ttg aaa aat tta gaa gaa tta aag aca gaa att gca aag 336 Leu Ser Lys Leu Lys Asn Leu Glu Glu Leu Lys Thr Glu Ile Ala Lys aag aaa tgt tcc gaa gaa ttt act aat aaa cta aaa agt ggt cat 384 Ala Lys Lys Cys Ser Glu Glu Phe Thr Asn Lys Leu Lys Ser Gly His gat ctt ggc aaa cag gat gct acc gat gat cat gca aaa gca gct 432 Ala Asp Leu Gly Lys Gln Asp Ala Thr Asp Asp His Ala Lys Ala Ala tta aaa aca cat gca act acc gat aaa ggt gct aaa gaa ttt aaa 48eu Lys Thr His Ala Thr Thr Asp Lys Gly Ala Lys Glu Phe Lys gat tta ttt gaa tca gta gaa ggt ttg tta aaa gca gct caa gta gca 528 Asp Leu Phe Glu Ser Val Glu Gly Leu Leu Lys Ala Ala Gln Val Ala act aat tca gtt aaa gaa ctt aca agt cct gtt gta gca gaa agt 576 Leu Thr Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser aaa aaa cct tcc atg gcc gtt tca gta gat ttg cct ggt gaa atg 624 Pro Lys Lys Pro Ser Met Ala Val Ser Val Asp Leu Pro Gly Glu Met 2gtt ctt gta agc aaa gaa aaa aac aaa gac ggc aag tac gat cta 672 Lys Val Leu Val Ser Lys Glu Lys Asn Lys Asp Gly Lys Tyr Asp Leu 222ca aca gta gac aag ctt gag ctt aaa gga act tct gat aaa aac 72la Thr Val Asp Lys Leu Glu Leu Lys Gly Thr Ser Asp Lys Asn 225 234ga tct gga gta ctt gaa ggc gta aaa gct gac aaa agt aaa gta 768 Asn Gly Ser Gly Val Leu Glu Gly Val Lys Ala Asp Lys Ser Lys Val 245 25aa tta aca att tct gac gat cta ggt caa acc aca ctt gaa gtt ttc 8Leu Thr Ile Ser Asp Asp Leu Gly Gln Thr Thr Leu Glu Val Phe 267aa gat ggc aaa aca cta gta tca aaa aaa gta act tcc aaa gac 864 Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Lys Val Thr Ser Lys Asp 275 28ag tca tca aca gaa gaa aaa ttc aat gaa aaa ggt gaa gta tct gaa 9Ser Ser Thr Glu Glu Lys Phe Asn Glu Lys Gly Glu Val Ser Glu 29ata ata aca aga gca gac gga acc aga ctt gaa tac aca gga att 96le Ile Thr Arg Ala Asp Gly Thr Arg Leu Glu Tyr Thr Gly Ile 33aaa agc gat gga tct gga aaa gct aaa gag gtt tta aaa aaa ttt act s Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys Lys Phe Thr 325 33tt gaa gga aaa gta gct aat gat aaa gta aca ttg gaa gta aaa gaa u Glu Gly Lys Val Ala Asn Asp Lys Val Thr Leu Glu Val Lys Glu 345cc gtt act tta agt aag aat att tca aaa tct ggg gaa gtt tca y Thr Val Thr Leu Ser Lys Asn Ile Ser Lys Ser Gly Glu Val Ser 355 36tt gaa ctt aat gac act gac agt agt gct gct act aaa aaa act gca l Glu Leu Asn Asp Thr Asp Ser Ser Ala Ala Thr Lys Lys Thr Ala 378gg aat tca aaa act tcc act tta aca att agt gtg aat agc caa a Trp Asn Ser Lys Thr Ser Thr Leu Thr Ile Ser Val Asn Ser Gln 385 39acc aaa aac ctt gta ttc aca aaa gaa gac aca ata aca gta caa s Thr Lys Asn Leu Val Phe Thr Lys Glu Asp Thr Ile Thr Val Gln 44tac gac tca gca ggc acc aat cta gaa ggc aaa gca gtc gaa att s Tyr Asp Ser Ala Gly Thr Asn Leu Glu Gly Lys Ala Val Glu Ile 423ca ctt aaa gaa ctt aaa aac gct tta aaa taa r Thr Leu Lys Glu Leu Lys Asn Ala Leu Lys * 435 4443 PRT Artificial Sequence Chimeric protein Ala Cys Ser Asn Ser Gly Lys Gly Gly Asp Ser Ala Ser Thr Asn Ala Asp Glu Ser Ala Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys 2 Lys Ile Thr Asp Ser Asn Ala Phe Val Leu Ala Val Lys Glu Val Glu 35 4r Leu Val Leu Ser Ile Asp Glu Leu Ala Lys Lys Ala Ile Gly Gln 5 Lys Ile Asp Asn Asn Asn Gly Leu Ala Ala Leu Asn Asn Gln Asn Gly 65 7 Ser Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Thr Glu Lys 85 9u Ser Lys Leu Lys Asn Leu Glu Glu Leu Lys Thr Glu Ile Ala Lys Lys Lys Cys Ser Glu Glu Phe Thr Asn Lys Leu Lys Ser Gly His Asp Leu Gly Lys Gln Asp Ala Thr Asp Asp His Ala Lys Ala Ala Leu Lys Thr His Ala Thr Thr Asp Lys Gly Ala Lys Glu Phe Lys Asp Leu Phe Glu Ser Val Glu Gly Leu Leu Lys Ala Ala Gln Val Ala Thr Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Lys Lys Pro Ser Met Ala Val Ser Val Asp Leu Pro Gly Glu Met 2Val Leu Val Ser Lys Glu Lys Asn Lys Asp Gly Lys Tyr Asp Leu 222la Thr Val Asp Lys Leu Glu Leu Lys Gly Thr Ser Asp Lys Asn 225 234ly Ser Gly Val Leu Glu Gly Val Lys Ala Asp Lys Ser Lys Val 245 25ys Leu Thr Ile Ser Asp Asp Leu Gly Gln Thr Thr Leu Glu Val Phe 267lu Asp Gly Lys Thr Leu Val Ser Lys Lys Val Thr Ser Lys Asp 275 28ys Ser Ser Thr Glu Glu Lys Phe Asn Glu Lys Gly Glu Val Ser Glu 29Ile Ile Thr Arg Ala Asp Gly Thr Arg Leu Glu Tyr Thr Gly Ile 33Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys Lys Phe Thr 325 33eu Glu Gly Lys Val Ala Asn Asp Lys Val Thr Leu Glu Val Lys Glu 345hr Val Thr Leu Ser Lys Asn Ile Ser Lys Ser Gly Glu Val Ser 355 36al Glu Leu Asn Asp Thr Asp Ser Ser Ala Ala Thr Lys Lys Thr Ala 378rp Asn Ser Lys Thr Ser Thr Leu Thr Ile Ser Val Asn Ser Gln 385 39Thr Lys Asn Leu Val Phe Thr Lys Glu Asp Thr Ile Thr Val Gln 44Tyr Asp Ser Ala Gly Thr Asn Leu Glu Gly Lys Ala Val Glu Ile 423hr Leu Lys Glu Leu Lys Asn Ala Leu Lys 435 443Artificial Sequence CDS (3meric nucleic acid gct tgt aat aat tca ggt ggg gat tct gca tct act aat cct gat 48 Met Ala Cys Asn Asn Ser Gly Gly Asp Ser Ala Ser Thr Asn Pro Asp tct gca aaa gga cct aat ctt acc gta ata agc aaa aaa att aca 96 Glu Ser Ala Lys Gly Pro Asn Leu Thr Val Ile Ser Lys Lys Ile Thr 2 gat tct aat gca ttt tta ctg gct gtg aaa gaa gtt gag gct ttg ctt Ser Asn Ala Phe Leu Leu Ala Val Lys Glu Val Glu Ala Leu Leu 35 4a tct ata gat gaa ctt tct aaa gct att ggt aaa aaa ata aaa aat Ser Ile Asp Glu Leu Ser Lys Ala Ile Gly Lys Lys Ile Lys Asn 5 gat ggt act tta gat aac gaa gca aat cga aac gaa tca ttg ata gca 24ly Thr Leu Asp Asn Glu Ala Asn Arg Asn Glu Ser Leu Ile Ala 65 7 gga gct tat gaa ata tca aaa cta ata aca caa aaa tta agt gta ttg 288 Gly Ala Tyr Glu Ile Ser Lys Leu Ile Thr Gln Lys Leu Ser Val Leu 85 9t tca gaa gaa tta aag gaa aaa att aaa gag gct aag gat tgt tcc 336 Asn Ser Glu Glu Leu Lys Glu Lys Ile Lys Glu Ala Lys Asp Cys Ser aaa ttt act act aag cta aaa gat agt cat gca gag ctt ggt ata 384 Glu Lys Phe Thr Thr Lys Leu Lys Asp Ser His Ala Glu Leu Gly Ile agc gtt cag gat gat aat gca aaa aaa gct att tta aaa aca cat 432 Gln Ser Val Gln Asp Asp Asn Ala Lys Lys Ala Ile Leu Lys Thr His act aaa gac aag ggt gct aaa gaa ctt gaa gag tta ttt aaa tca 48hr Lys Asp Lys Gly Ala Lys Glu Leu Glu Glu Leu Phe Lys Ser cta gaa agc ttg tca aaa gca gcg caa gca gca tta act aat tca gtt 528 Leu Glu Ser Leu Ser Lys Ala Ala Gln Ala Ala Leu Thr Asn Ser Val gag ctt aca aat cct gtt gtg gca gaa agt cca aaa aaa cct tcc 576 Lys Glu Leu Thr Asn Pro Val Val Ala Glu Ser Pro Lys Lys Pro Ser gcc gtt tca gta gat ttg cct ggt gaa atg aaa gtt ctt gta agc 624 Met Ala Val Ser Val Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser 2gaa aaa aac aaa gac ggc aag tac gat cta att gca aca gta gac 672 Lys Glu Lys Asn Lys Asp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp 222tt gag ctt aaa gga act tct gat aaa aac aat gga tct gga gta 72eu Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val 225 234aa ggc gta aaa gct gac aaa agt aaa gta aaa tta aca att tct 768 Leu Glu Gly Val Lys Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser 245 25ac gat cta ggt caa acc aca ctt gaa gtt ttc aaa gaa gat ggc aaa 8Asp Leu Gly Gln Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys 267ta gta tca aaa aaa gta act tcc aaa gac aag tca tca aca gaa 864 Thr Leu Val Ser Lys Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu 275 28aa aaa ttc aat gaa aaa ggt gaa gta tct gaa aaa ata ata aca aga 9Lys Phe Asn Glu Lys Gly Glu Val Ser Glu Lys Ile Ile Thr

Arg 29gac gga acc aga ctt gaa tac aca gga att aaa agc gat gga tct 96sp Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser 33gga aaa gct aaa gag gtt tta aaa aaa ttt act ctt gaa gga aaa gta y Lys Ala Lys Glu Val Leu Lys Lys Phe Thr Leu Glu Gly Lys Val 325 33ct aat gat aaa gta aca ttg gaa gta aaa gaa gga acc gtt act tta a Asn Asp Lys Val Thr Leu Glu Val Lys Glu Gly Thr Val Thr Leu 345ag aac att tca aaa tct ggg gaa gtt tca gtt gaa ctt aat gac r Lys Asn Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp 355 36ct gac agt agt gct gct act aaa aaa act gca gct tgg aat tca aaa r Asp Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys 378ct act tta aca att agt gtt aac agc aaa aaa act aca caa ctt r Ser Thr Leu Thr Ile Ser Val Asn Ser Lys Lys Thr Thr Gln Leu 385 39ttt act aaa caa gac aca ata act gta caa aaa tac gac tcc gca l Phe Thr Lys Gln Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala 44acc aat tta gaa ggc aca gca gtc gaa att aaa aca ctt gat gaa y Thr Asn Leu Glu Gly Thr Ala Val Glu Ile Lys Thr Leu Asp Glu 423aa aac gct tta aaa taa u Lys Asn Ala Leu Lys * 435 PRT Artificial Sequence Chimeric protein Ala Cys Asn Asn Ser Gly Gly Asp Ser Ala Ser Thr Asn Pro Asp Ser Ala Lys Gly Pro Asn Leu Thr Val Ile Ser Lys Lys Ile Thr 2 Asp Ser Asn Ala Phe Leu Leu Ala Val Lys Glu Val Glu Ala Leu Leu 35 4r Ser Ile Asp Glu Leu Ser Lys Ala Ile Gly Lys Lys Ile Lys Asn 5 Asp Gly Thr Leu Asp Asn Glu Ala Asn Arg Asn Glu Ser Leu Ile Ala 65 7 Gly Ala Tyr Glu Ile Ser Lys Leu Ile Thr Gln Lys Leu Ser Val Leu 85 9n Ser Glu Glu Leu Lys Glu Lys Ile Lys Glu Ala Lys Asp Cys Ser Lys Phe Thr Thr Lys Leu Lys Asp Ser His Ala Glu Leu Gly Ile Ser Val Gln Asp Asp Asn Ala Lys Lys Ala Ile Leu Lys Thr His Thr Lys Asp Lys Gly Ala Lys Glu Leu Glu Glu Leu Phe Lys Ser Leu Glu Ser Leu Ser Lys Ala Ala Gln Ala Ala Leu Thr Asn Ser Val Glu Leu Thr Asn Pro Val Val Ala Glu Ser Pro Lys Lys Pro Ser Ala Val Ser Val Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser 2Glu Lys Asn Lys Asp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp 222eu Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val 225 234lu Gly Val Lys Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser 245 25sp Asp Leu Gly Gln Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys 267eu Val Ser Lys Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu 275 28lu Lys Phe Asn Glu Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg 29Asp Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser 33Gly Lys Ala Lys Glu Val Leu Lys Lys Phe Thr Leu Glu Gly Lys Val 325 33la Asn Asp Lys Val Thr Leu Glu Val Lys Glu Gly Thr Val Thr Leu 345ys Asn Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp 355 36hr Asp Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys 378er Thr Leu Thr Ile Ser Val Asn Ser Lys Lys Thr Thr Gln Leu 385 39Phe Thr Lys Gln Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala 44Thr Asn Leu Glu Gly Thr Ala Val Glu Ile Lys Thr Leu Asp Glu 423ys Asn Ala Leu Lys 435 9 DNA Artificial Sequence CDS (imeric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata aat aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gaa att gct gct aaa gct att ggt aaa aaa Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 ata cac caa aat aat ggt ttg gat acc gaa aat aat cac aat gga tca 24is Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 ttg tta gcg gga gct tat gca ata tca acc cta ata aaa caa aaa tta 288 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9t gga ttg aaa aat gaa gga tta aag gaa aaa att gat gcg gct aag 336 Asp Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys tgt tct gaa aca ttt act aat aaa tta aaa gaa aaa cac aca gat 384 Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp ggt aaa gaa ggt gtt act gat gct gat gca aaa gaa gcc att tta 432 Leu Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu gca aat ggt act aaa act aaa ggt gct gaa gaa ctt gga aaa tta 48la Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu ttt gaa tca gta gag gtc ttg tca aaa gca gct aaa gag atg ctt gct 528 Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala tca gtt aaa gag ctt aca agc cct gtt gtg gca gaa agt cca aaa 576 Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys cct tcc atg gcc aag caa aat gtt tct gaa aaa ata ata aca aga 624 Lys Pro Ser Met Ala Lys Gln Asn Val Ser Glu Lys Ile Ile Thr Arg 2gac gga acc aga ctt gaa tac aca gga att aaa agc gat gga tct 672 Ala Asp Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser 222aa gct aaa gag gtt tta aaa ggc tat gtt ctt gaa gga act cta 72ys Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu 225 234ct gaa aaa aca aca ttg gtg gtt aaa gaa gga act gtt act tta 768 Thr Ala Glu Lys Thr Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu 245 25gc aaa aat att tca aaa tct ggg gaa gtt tca gtt gaa ctt aat gac 8Lys Asn Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp 267ac agt agt gct gct act aaa aaa act gca gct tgg aat tca ggc 864 Thr Asp Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Gly 275 28ct tca act tta aca att act gta aac agt aaa aaa act aaa gac ctt 9Ser Thr Leu Thr Ile Thr Val Asn Ser Lys Lys Thr Lys Asp Leu 29ttt aca aaa gaa aac aca att aca gta caa caa tac gac tca aat 96he Thr Lys Glu Asn Thr Ile Thr Val Gln Gln Tyr Asp Ser Asn 33ggc acc aaa tta gag ggg tca gca gtt gaa att aca aaa ctt gat gaa y Thr Lys Leu Glu Gly Ser Ala Val Glu Ile Thr Lys Leu Asp Glu 325 33tt aaa aac gct tta aaa taa e Lys Asn Ala Leu Lys * 3442 PRT Artificial Sequence Chimeric protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 Ile His Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9p Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu Ala Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys Pro Ser Met Ala Lys Gln Asn Val Ser Glu Lys Ile Ile Thr Arg 2Asp Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser 222ys Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu 225 234la Glu Lys Thr Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu 245 25er Lys Asn Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp 267sp Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Gly 275 28hr Ser Thr Leu Thr Ile Thr Val Asn Ser Lys Lys Thr Lys Asp Leu 29Phe Thr Lys Glu Asn Thr Ile Thr Val Gln Gln Tyr Asp Ser Asn 33Gly Thr Lys Leu Glu Gly Ser Ala Val Glu Ile Thr Lys Leu Asp Glu 325 33le Lys Asn Ala Leu Lys 34 Artificial Sequence CDS (imeric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata aat aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gaa att gct gct aaa gct att ggt aaa aaa Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 ata cac caa aat aat ggt ttg gat acc gaa aat aat cac aat gga tca 24is Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 ttg tta gcg gga gct tat gca ata tca acc cta ata aaa caa aaa tta 288 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9t gga ttg aaa aat gaa gga tta aag gaa aaa att gat gcg gct aag 336 Asp Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys tgt tct gaa aca ttt act aat aaa tta aaa gaa aaa cac aca gat 384 Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp ggt aaa gaa ggt gtt act gat gct gat gca aaa gaa gcc att tta 432 Leu Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu gca aat ggt act aaa act aaa ggt gct gaa gaa ctt gga aaa tta 48la Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu ttt gaa tca gta gag gtc ttg tca aaa gca gct aaa gag atg ctt gct 528 Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala tca gtt aaa gag ctt aca agc cct gtt gtg gca gaa agt cca aaa 576 Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys cct tcc atg gcc aag caa aat gtt tct gaa aaa ata ata aca aga 624 Lys Pro Ser Met Ala Lys Gln Asn Val Ser Glu Lys Ile Ile Thr Arg 2gac gga acc aga ctt gaa tac aca gga att aaa agc gat gga tct 672 Ala Asp Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser 222aa gct aaa gag gtt tta aaa ggc tat gtt ctt gaa gga act cta 72ys Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu 225 234ct gaa aaa aca aca ttg gtg gtt aaa gaa gga act gtt act tta 768 Thr Ala Glu Lys Thr Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu 245 25gc aaa aat att tca aaa tct ggg gaa gtt tca gtt gaa ctt aat gac 8Lys Asn Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp 267ac agt agt gct gct act aaa aaa act gca gct tgg aat tca aaa 864 Thr Asp Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys 275 28ct tcc act tta aca att agt gtg aat agc caa aaa acc aaa aac ctt 9Ser Thr Leu Thr Ile Ser Val Asn Ser Gln Lys Thr Lys Asn Leu 29ttc aca aaa gaa gac aca ata aca gta caa aaa tac gac tca gca 96he Thr Lys Glu Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala 33ggc acc aat cta gaa ggc aaa gca gtc gaa att aca aca ctt aaa gaa y Thr Asn Leu Glu Gly Lys Ala Val Glu Ile Thr Thr Leu Lys Glu 325 33tt aaa aac gct tta aaa taa u Lys Asn Ala Leu Lys * 3442 PRT Artificial Sequence Chimeric protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 Ile His Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9p Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu Ala Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys Pro Ser Met Ala Lys Gln Asn Val Ser Glu Lys Ile Ile Thr Arg 2Asp Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser 222ys Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu 225 234la Glu Lys Thr Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu 245 25er Lys Asn Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp 267sp Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys 275 28BR> 285 Thr Ser Thr Leu Thr Ile Ser Val Asn Ser Gln Lys Thr Lys Asn Leu 29Phe Thr Lys Glu Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala 33Gly Thr Asn Leu Glu Gly Lys Ala Val Glu Ile Thr Thr Leu Lys Glu 325 33eu Lys Asn Ala Leu Lys 34 Artificial Sequence CDS (imeric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata aat aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gaa att gct gct aaa gct att ggt aaa aaa Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 ata cac caa aat aat ggt ttg gat acc gaa aat aat cac aat gga tca 24is Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 ttg tta gcg gga gct tat gca ata tca acc cta ata aaa caa aaa tta 288 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9t gga ttg aaa aat gaa gga tta aag gaa aaa att gat gcg gct aag 336 Asp Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys tgt tct gaa aca ttt act aat aaa tta aaa gaa aaa cac aca gat 384 Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp ggt aaa gaa ggt gtt act gat gct gat gca aaa gaa gcc att tta 432 Leu Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu gca aat ggt act aaa act aaa ggt gct gaa gaa ctt gga aaa tta 48la Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu ttt gaa tca gta gag gtc ttg tca aaa gca gct aaa gag atg ctt gct 528 Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala tca gtt aaa gag ctt aca agc cct gtt gtg gca gaa agt cca aaa 576 Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys cct tcc atg gcc aag caa aat gtt tct gaa aaa ata ata aca aga 624 Lys Pro Ser Met Ala Lys Gln Asn Val Ser Glu Lys Ile Ile Thr Arg 2gac gga acc aga ctt gaa tac aca gga att aaa agc gat gga tct 672 Ala Asp Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser 222aa gct aaa gag gtt tta aaa ggc tat gtt ctt gaa gga act cta 72ys Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu 225 234ct gaa aaa aca aca ttg gtg gtt aaa gaa gga act gtt act tta 768 Thr Ala Glu Lys Thr Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu 245 25gc aaa aat att tca aaa tct ggg gaa gtt tca gtt gaa ctt aat gac 8Lys Asn Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp 267ac agt agt gct gct act aaa aaa act gca gct tgg aat tca aaa 864 Thr Asp Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys 275 28ct tct act tta aca att agt gtt aac agc aaa aaa act aca caa ctt 9Ser Thr Leu Thr Ile Ser Val Asn Ser Lys Lys Thr Thr Gln Leu 29ttt act aaa caa gac aca ata act gta caa aaa tac gac tcc gca 96he Thr Lys Gln Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala 33ggt acc aat tta gaa ggc aca gca gtc gaa att aaa aca ctt gat gaa y Thr Asn Leu Glu Gly Thr Ala Val Glu Ile Lys Thr Leu Asp Glu 325 33tt aaa aac gct tta aaa taa u Lys Asn Ala Leu Lys * 3442 PRT Artificial Sequence Chimeric protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 Ile His Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9p Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu Ala Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys Pro Ser Met Ala Lys Gln Asn Val Ser Glu Lys Ile Ile Thr Arg 2Asp Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser 222ys Ala Lys Glu Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu 225 234la Glu Lys Thr Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu 245 25er Lys Asn Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp 267sp Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys 275 28hr Ser Thr Leu Thr Ile Ser Val Asn Ser Lys Lys Thr Thr Gln Leu 29Phe Thr Lys Gln Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala 33Gly Thr Asn Leu Glu Gly Thr Ala Val Glu Ile Lys Thr Leu Asp Glu 325 33eu Lys Asn Ala Leu Lys 34 Artificial Sequence CDS (imeric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata agt aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gaa att gct gct aaa gct att ggt aaa aaa Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 ata cac caa aat aat ggt ttg gat acc gaa tat aat cac aat gga tca 24is Gln Asn Asn Gly Leu Asp Thr Glu Tyr Asn His Asn Gly Ser 65 7 ttg tta gcg gga gct tat gca ata tca acc cta ata aaa caa aaa tta 288 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9t gga ttg aaa aat gaa gga tta aag gaa aaa att gat gcg gct aag 336 Asp Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys tgt tct gaa aca ttt act aat aaa tta aaa gaa aaa cac aca gat 384 Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp ggt aaa gaa ggt gtt act gat gct gat gca aaa gaa gcc att tta 432 Leu Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu aca aat ggt act aaa act aaa ggt gct gaa gaa ctt gga aaa tta 48hr Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu ttt gaa tca gta gag gtc ttg tca aaa gca gct aaa gag atg ctt gct 528 Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala tca gtt aaa gag ctt aca agc cct gtt gtg gca gaa agt cca aaa 576 Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys cct tcc atg gcc aag caa aat gtt aca tct gaa aaa aca ata gta 624 Lys Pro Ser Met Ala Lys Gln Asn Val Thr Ser Glu Lys Thr Ile Val 2gca aat gga acc aga ctt gaa tac aca gac ata aaa agc gat gga 672 Arg Ala Asn Gly Thr Arg Leu Glu Tyr Thr Asp Ile Lys Ser Asp Gly 222ga aaa gct aaa gaa gtt tta aaa gac ttt act ctt gaa gga act 72ly Lys Ala Lys Glu Val Leu Lys Asp Phe Thr Leu Glu Gly Thr 225 234ct gct gac ggc aaa aca aca ttg aaa gtt aca gaa ggc act gtt 768 Leu Ala Ala Asp Gly Lys Thr Thr Leu Lys Val Thr Glu Gly Thr Val 245 25tt tta agc aag aac att tta aaa tcc gga gaa ata aca gtt gca ctt 8Leu Ser Lys Asn Ile Leu Lys Ser Gly Glu Ile Thr Val Ala Leu 267ac tct gac act act cag gct act aaa aaa act gga aaa tgg gat 864 Asp Asp Ser Asp Thr Thr Gln Ala Thr Lys Lys Thr Gly Lys Trp Asp 275 28ca aat act tcc act tta aca att agt gtg aat agc aaa aaa act aaa 9Asn Thr Ser Thr Leu Thr Ile Ser Val Asn Ser Lys Lys Thr Lys 29att gta ttt aca aaa gaa gac aca ata aca gta caa aaa tac gac 96le Val Phe Thr Lys Glu Asp Thr Ile Thr Val Gln Lys Tyr Asp 33tca gca ggc acc aat cta gaa ggc aac gca gtc gaa att aaa aca ctt r Ala Gly Thr Asn Leu Glu Gly Asn Ala Val Glu Ile Lys Thr Leu 325 33at gaa ctt aaa aac gct tta aaa tag p Glu Leu Lys Asn Ala Leu Lys * 3444 PRT Artificial Sequence Chimeric protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 Ile His Gln Asn Asn Gly Leu Asp Thr Glu Tyr Asn His Asn Gly Ser 65 7 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9p Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu Thr Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys Pro Ser Met Ala Lys Gln Asn Val Thr Ser Glu Lys Thr Ile Val 2Ala Asn Gly Thr Arg Leu Glu Tyr Thr Asp Ile Lys Ser Asp Gly 222ly Lys Ala Lys Glu Val Leu Lys Asp Phe Thr Leu Glu Gly Thr 225 234la Ala Asp Gly Lys Thr Thr Leu Lys Val Thr Glu Gly Thr Val 245 25al Leu Ser Lys Asn Ile Leu Lys Ser Gly Glu Ile Thr Val Ala Leu 267sp Ser Asp Thr Thr Gln Ala Thr Lys Lys Thr Gly Lys Trp Asp 275 28er Asn Thr Ser Thr Leu Thr Ile Ser Val Asn Ser Lys Lys Thr Lys 29Ile Val Phe Thr Lys Glu Asp Thr Ile Thr Val Gln Lys Tyr Asp 33Ser Ala Gly Thr Asn Leu Glu Gly Asn Ala Val Glu Ile Lys Thr Leu 325 33sp Glu Leu Lys Asn Ala Leu Lys 34323 DNA Artificial Sequence CDS (323) Chimeric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata aat aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gaa att gct gct aaa gct att ggt aaa aaa Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 ata cac caa aat aat ggt ttg gat acc gaa aat aat cac aat gga tca 24is Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 ttg tta gcg gga gct tat gca ata tca acc cta ata aaa caa aaa tta 288 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9t gga ttg aaa aat gaa gga tta aag gaa aaa att gat gcg gct aag 336 Asp Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys tgt tct gaa aca ttt act aat aaa tta aaa gaa aaa cac aca gat 384 Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp ggt aaa gaa ggt gtt act gat gct gat gca aaa gaa gcc att tta 432 Leu Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu gca aat ggt act aaa act aaa ggt gct gaa gaa ctt gga aaa tta 48la Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu ttt gaa tca gta gag gtc ttg tca aaa gca gct aaa gag atg ctt gct 528 Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala tca gtt aaa gag ctt aca agc cct gtt gtg gca gaa agt cca aaa 576 Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys cct tcc atg gcc gtt tca gta gat ttg cct ggt gaa atg aaa gtt 624 Lys Pro Ser Met Ala Val Ser Val Asp Leu Pro Gly Glu Met Lys Val 2gta agc aaa gaa aaa aac aaa gac ggc aag tac gat cta att gca 672 Leu Val Ser Lys Glu Lys Asn Lys Asp Gly Lys Tyr Asp Leu Ile Ala 222ta gac aag ctt gag ctt aaa gga act tct gat aaa aac aat gga 72al Asp Lys Leu Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn Gly 225 234ga gta ctt gaa ggc gta aaa gct gac aaa agt aaa gta aaa tta 768 Ser Gly Val Leu Glu Gly Val Lys Ala Asp Lys Ser Lys Val Lys Leu 245 25ca att tct gac gat cta ggt caa acc aca ctt gaa gtt ttc aaa gaa 8Ile Ser Asp Asp Leu Gly Gln Thr Thr Leu Glu Val Phe Lys Glu 267gc aaa aca cta gta tca aaa aaa gta act tcc aaa gac aag tca 864 Asp Gly Lys Thr Leu Val Ser Lys Lys Val Thr Ser Lys Asp Lys Ser 275 28ca aca gaa gaa aaa ttc aat gaa aaa ggt gaa gta tct gaa aaa ata 9Thr Glu Glu Lys Phe Asn Glu Lys Gly Glu Val Ser Glu Lys Ile 29aca aga gca gac gga acc aga ctt gaa tac aca gga att aaa agc 96hr Arg Ala Asp Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser 33gat gga tct gga aaa gct aaa gag gtt tta aaa ggc ttt act ctt gaa p Gly Ser Gly Lys Ala Lys Glu Val Leu Lys Gly Phe Thr Leu Glu 325 33ga aaa gta gct aat gat aaa gta aca ttg gaa gta aaa gaa gga acc y

Lys Val Ala Asn Asp Lys Val Thr Leu Glu Val Lys Glu Gly Thr 345ct tta agt aag att tca aaa tct ggg gaa gtt tca gtt gaa ctt l Thr Leu Ser Lys Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu 355 36at gac act gac agt agt gct gct act aaa aaa act gca gct tgg aat n Asp Thr Asp Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn 378aa act tct act tta aca att agt gtt aac agc aaa aaa act aca r Lys Thr Ser Thr Leu Thr Ile Ser Val Asn Ser Lys Lys Thr Thr 385 39ctt gtg ttt act aaa caa gac aca ata act gta caa aaa tac gac n Leu Val Phe Thr Lys Gln Asp Thr Ile Thr Val Gln Lys Tyr Asp 44gca ggt acc aat tta gaa ggc aca gca gtc gaa att aaa aca ctt r Ala Gly Thr Asn Leu Glu Gly Thr Ala Val Glu Ile Lys Thr Leu 423aa ctt aaa aac gct tta aaa taa p Glu Leu Lys Asn Ala Leu Lys * 435 444rtificial Sequence Chimeric protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Asn Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Ile Ala Ala Lys Ala Ile Gly Lys Lys 5 Ile His Gln Asn Asn Gly Leu Asp Thr Glu Asn Asn His Asn Gly Ser 65 7 Leu Leu Ala Gly Ala Tyr Ala Ile Ser Thr Leu Ile Lys Gln Lys Leu 85 9p Gly Leu Lys Asn Glu Gly Leu Lys Glu Lys Ile Asp Ala Ala Lys Cys Ser Glu Thr Phe Thr Asn Lys Leu Lys Glu Lys His Thr Asp Gly Lys Glu Gly Val Thr Asp Ala Asp Ala Lys Glu Ala Ile Leu Ala Asn Gly Thr Lys Thr Lys Gly Ala Glu Glu Leu Gly Lys Leu Phe Glu Ser Val Glu Val Leu Ser Lys Ala Ala Lys Glu Met Leu Ala Ser Val Lys Glu Leu Thr Ser Pro Val Val Ala Glu Ser Pro Lys Pro Ser Met Ala Val Ser Val Asp Leu Pro Gly Glu Met Lys Val 2Val Ser Lys Glu Lys Asn Lys Asp Gly Lys Tyr Asp Leu Ile Ala 222al Asp Lys Leu Glu Leu Lys Gly Thr Ser Asp Lys Asn Asn Gly 225 234ly Val Leu Glu Gly Val Lys Ala Asp Lys Ser Lys Val Lys Leu 245 25hr Ile Ser Asp Asp Leu Gly Gln Thr Thr Leu Glu Val Phe Lys Glu 267ly Lys Thr Leu Val Ser Lys Lys Val Thr Ser Lys Asp Lys Ser 275 28er Thr Glu Glu Lys Phe Asn Glu Lys Gly Glu Val Ser Glu Lys Ile 29Thr Arg Ala Asp Gly Thr Arg Leu Glu Tyr Thr Gly Ile Lys Ser 33Asp Gly Ser Gly Lys Ala Lys Glu Val Leu Lys Gly Phe Thr Leu Glu 325 33ly Lys Val Ala Asn Asp Lys Val Thr Leu Glu Val Lys Glu Gly Thr 345hr Leu Ser Lys Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu 355 36sn Asp Thr Asp Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn 378ys Thr Ser Thr Leu Thr Ile Ser Val Asn Ser Lys Lys Thr Thr 385 39Leu Val Phe Thr Lys Gln Asp Thr Ile Thr Val Gln Lys Tyr Asp 44Ala Gly Thr Asn Leu Glu Gly Thr Ala Val Glu Ile Lys Thr Leu 423lu Leu Lys Asn Ala Leu Lys 435 443Artificial Sequence CDS (3meric nucleic acid gct tgt aat aat tca ggg aaa gat ggg aat aca tct gca aat tct 48 Met Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser gat gag tct gtt aaa ggg cct aat ctt aca gaa ata agt aaa aaa 96 Ala Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 att acg gat tct aat gcg gtt tta ctt gct gtg aaa gag gtt gaa gcg Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4g ctg tca tct ata gat gag ctt gct aaa gct att ggt aaa aaa ata Leu Ser Ser Ile Asp Glu Leu Ala Lys Ala Ile Gly Lys Lys Ile 5 aaa aac gat ggt agt tta gat aat gaa gca aat cgc aac gag tca ttg 24sn Asp Gly Ser Leu Asp Asn Glu Ala Asn Arg Asn Glu Ser Leu 65 7 tta gca gga gct tat aca ata tca acc tta ata aca caa aaa tta agt 288 Leu Ala Gly Ala Tyr Thr Ile Ser Thr Leu Ile Thr Gln Lys Leu Ser 85 9a tta aac gga tca gaa ggt tta aag gaa aag att gcc gca gct aag 336 Lys Leu Asn Gly Ser Glu Gly Leu Lys Glu Lys Ile Ala Ala Ala Lys tgc tct gaa gag ttt agt act aaa cta aaa gat aat cat gca cag 384 Lys Cys Ser Glu Glu Phe Ser Thr Lys Leu Lys Asp Asn His Ala Gln ggt ata cag ggc gtt act gat gaa aat gca aaa aaa gct att tta 432 Leu Gly Ile Gln Gly Val Thr Asp Glu Asn Ala Lys Lys Ala Ile Leu gca aat gca gcg ggt aaa gat aag ggc gtt gaa gaa ctt gaa aag 48la Asn Ala Ala Gly Lys Asp Lys Gly Val Glu Glu Leu Glu Lys ttg tcc gga tca tta gaa agc tta tca aaa gca gct aaa gag atg ctt 528 Leu Ser Gly Ser Leu Glu Ser Leu Ser Lys Ala Ala Lys Glu Met Leu aat tca gtt aaa gag ctt aca agc cct gtt gtc cat ggc gtt tca 576 Ala Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val His Gly Val Ser gat ttg cct ggt gaa atg aaa gtt ctt gta agc aaa gaa aaa aac 624 Val Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn 2gac ggc aag tac gat cta att gca aca gta gac aag ctt gag ctt 672 Lys Asp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu 222ga act tct gat aaa aac aat gga tct gga gta ctt gaa ggc gta 72ly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val 225 234ct gac aaa agt aaa gta aaa tta aca att tct gac gat cta ggt 768 Lys Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly 245 25aa acc aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca cta gta tca 8Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser 267aa gta act tcc aaa gac aag tca tca aca gaa gaa aaa ttc aat 864 Lys Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn 275 28aa aaa ggt gaa gta tct gaa aaa ata ata aca aga gca gac gga acc 9Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr 29ctt gaa tac aca gga att aaa agc gat gga tct gga aaa gct aaa 96eu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys 33gag gtt tta aaa ggc ttt act ctt gaa gga aaa gta gct aat gat aaa u Val Leu Lys Gly Phe Thr Leu Glu Gly Lys Val Ala Asn Asp Lys 325 33ta aca ttg gaa gta aaa gaa gga acc gtt act tta agt aag att tca l Thr Leu Glu Val Lys Glu Gly Thr Val Thr Leu Ser Lys Ile Ser 345ct ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt gct s Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 355 36ct act aaa aaa act gca gct tgg aat tca aaa act tct act tta aca a Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr 378gt gtt aac agc aaa aaa act aca caa ctt gtg ttt act aaa caa e Ser Val Asn Ser Lys Lys Thr Thr Gln Leu Val Phe Thr Lys Gln 385 39aca ata act gta caa aaa tac gac tcc gca ggt acc aat tta gaa p Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 44aca gca gtc gaa att aaa aca ctt gat gaa ctt aaa aac gct tta y Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu 423aa s * PRT Artificial Sequence Chimeric protein Ala Cys Asn Asn Ser Gly Lys Asp Gly Asn Thr Ser Ala Asn Ser Asp Glu Ser Val Lys Gly Pro Asn Leu Thr Glu Ile Ser Lys Lys 2 Ile Thr Asp Ser Asn Ala Val Leu Leu Ala Val Lys Glu Val Glu Ala 35 4u Leu Ser Ser Ile Asp Glu Leu Ala Lys Ala Ile Gly Lys Lys Ile 5 Lys Asn Asp Gly Ser Leu Asp Asn Glu Ala Asn Arg Asn Glu Ser Leu 65 7 Leu Ala Gly Ala Tyr Thr Ile Ser Thr Leu Ile Thr Gln Lys Leu Ser 85 9s Leu Asn Gly Ser Glu Gly Leu Lys Glu Lys Ile Ala Ala Ala Lys Cys Ser Glu Glu Phe Ser Thr Lys Leu Lys Asp Asn His Ala Gln Gly Ile Gln Gly Val Thr Asp Glu Asn Ala Lys Lys Ala Ile Leu Ala Asn Ala Ala Gly Lys Asp Lys Gly Val Glu Glu Leu Glu Lys Leu Ser Gly Ser Leu Glu Ser Leu Ser Lys Ala Ala Lys Glu Met Leu Asn Ser Val Lys Glu Leu Thr Ser Pro Val Val His Gly Val Ser Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn 2Asp Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu 222ly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val 225 234la Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly 245 25ln Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser 267ys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn 275 28lu Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr 29Leu Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys 33Glu Val Leu Lys Gly Phe Thr Leu Glu Gly Lys Val Ala Asn Asp Lys 325 33al Thr Leu Glu Val Lys Glu Gly Thr Val Thr Leu Ser Lys Ile Ser 345er Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 355 36la Thr Lys Lys Thr Ala Ala Trp Asn Ser Lys Thr Ser Thr Leu Thr 378er Val Asn Ser Lys Lys Thr Thr Gln Leu Val Phe Thr Lys Gln 385 39Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 44Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu 4238A Artificial Sequence Oligonucleotide Primer atatggc ttgtaataat tcagggaaag a 3rtificial Sequence Oligonucleotide Primer ccatgga aggttttttt ggactttctg 3rtificial Sequence Oligonucleotide Primer ccatggc caagcaaaat gttagcagcc 37 DNA Artificial Sequence Oligonucleotide Primer ggatcct tattttaaag cgttttt 27 DNA Borrelia burgdorferi CDS ( atg aaa aaa tat tta ttg gga ata ggt cta ata tta gcc tta ata gca 48 Met Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala aag caa aat gtt agc agc ctt gac gag aaa aac agc gtt tca gta 96 Cys Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 gat ttg cct ggt gaa atg aaa gtt ctt gta agc aaa gaa aaa aac aaa Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys 35 4c ggc aag tac gat cta att gca aca gta gac aag ctt gag ctt aaa Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys 5 gga act tct gat aaa aac aat gga tct gga gta ctt gaa ggc gta aaa 24hr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 gct gac aaa agt aaa gta aaa tta aca att tct gac gat cta ggt caa 288 Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln 85 9c aca ctt gaa gtt ttc aaa gaa gat ggc aaa aca cta gta tca aaa 336 Thr Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys gta act tcc aaa gac aag tca tca aca gaa gaa aaa ttc aat gaa 384 Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu ggt gaa gta tct gaa aaa ata ata aca aga gca gac gga acc aga 432 Lys Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg gaa tac aca gga att aaa agc gat gga tct gga aaa gct aaa gag 48lu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu gtt tta aaa ggc tat gtt ctt gaa gga act cta act gct gaa aaa aca 528 Val Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr ttg gtg gtt aaa gaa gga act gtt act tta agc aaa aat att tca 576 Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser tct ggg gaa gtt tca gtt gaa ctt aat gac act gac agt agt gct 624 Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2act aaa aaa act gca gct tgg aat gca ggc act tca act tta aca 672 Ala Thr Lys Lys Thr Ala Ala Trp Asn Ala Gly Thr Ser Thr Leu Thr 222ct gta aac aac aaa aaa act aaa gcc ctt gta ttt aca aaa caa 72hr Val Asn Asn Lys Lys Thr Lys Ala Leu Val Phe Thr Lys Gln 225 234ca att aca tca caa aaa tac gac tca gca gga acc aac ttg gaa 768 Asp Thr Ile Thr Ser Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25gc aca gca gtc gaa att aaa aca ctt gat gaa ctt aaa aac gct tta 8Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu 267 PRT Borrelia burgdorferi Lys Lys Tyr Leu Leu Gly Ile Gly Leu Ile Leu Ala Leu Ile Ala Lys Gln Asn Val Ser Ser Leu Asp Glu Lys Asn Ser Val Ser Val 2 Asp Leu Pro Gly Glu Met Lys Val Leu Val Ser Lys Glu Lys Asn Lys 35 4p Gly Lys Tyr Asp Leu Ile Ala Thr Val Asp Lys Leu Glu Leu Lys 5 Gly Thr Ser Asp Lys Asn Asn Gly Ser Gly Val Leu Glu Gly Val Lys 65 7 Ala Asp Lys Ser Lys Val Lys Leu Thr Ile Ser Asp Asp Leu Gly Gln 85 9r Thr Leu Glu Val Phe Lys Glu Asp Gly Lys Thr Leu Val Ser Lys Val Thr Ser Lys Asp Lys Ser Ser Thr Glu Glu Lys Phe Asn Glu Gly Glu Val Ser Glu Lys Ile Ile Thr Arg Ala Asp Gly Thr Arg Glu Tyr Thr Gly Ile Lys Ser Asp Gly Ser Gly Lys Ala Lys Glu >
Leu Lys Gly Tyr Val Leu Glu Gly Thr Leu Thr Ala Glu Lys Thr Leu Val Val Lys Glu Gly Thr Val Thr Leu Ser Lys Asn Ile Ser Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ser Ala 2Thr Lys Lys Thr Ala Ala Trp Asn Ala Gly Thr Ser Thr Leu Thr 222hr Val Asn Asn Lys Lys Thr Lys Ala Leu Val Phe Thr Lys Gln 225 234hr Ile Thr Ser Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu 245 25ly Thr Ala Val Glu Ile Lys Thr Leu Asp Glu Leu Lys Asn Ala Leu 26787 8 PRT Borrelia burgdorferi sensu lato Pro Gly Glu Met Lys Val Leu 8 PRT Borrelia burgdorferi sensu lato Pro Gly Gly Met Thr Val Leu 8 PRT Borrelia burgdorferi sensu lato Pro Gly Glu Ile Lys Val Leu 8 PRT Borrelia burgdorferi sensu lato Pro Gly Gly Met Gly Val Leu Borrelia burgdorferi sensu lato Thr Ser Asp Lys Asn Asn Gly Ser Gly Val PRT Borrelia burgdorferi sensu lato Thr Ser Asp Lys Ser Asn Gly Ser Gly Thr PRT Borrelia burgdorferi sensu lato Thr Ser Asp Lys Asn Asn Gly Ser Gly Thr PRT Borrelia burgdorferi sensu lato Thr Ser Asp Lys Asp Asn Gly Ser Gly Thr PRT Borrelia burgdorferi sensu lato Thr Ser Asp Lys Asp Asn Gly Ser Gly Val PRT Borrelia burgdorferi sensu lato Ile Ser Lys Ser Gly Glu Val Ser Val Glu Leu Asn Asp Thr Asp Ser Ala Ala Thr Lys Lys Thr Ala Ala Trp Asn Ser Gly Thr 2 PRT Borrelia burgdorferi sensu lato Ile Pro Asn Ser Gly Glu Ile Thr Val Glu Leu Asn Asp Ser Asn Thr Gln Ala Thr Lys Lys Thr Gly Lys Trp Asp Ser Asn Thr 2 PRT Borrelia burgdorferi sensu lato Ile Leu Lys Ser Gly Glu Ile Thr Val Ala Leu Asp Asp Ser Asp Thr Gln Ala Thr Lys Lys Thr Gly Lys Trp Asp Ser Lys Thr 2 PRT Borrelia burgdorferi sensu lato Ile Leu Lys Ser Gly Glu Ile Thr Ala Ala Leu Asp Asp Ser Asp Thr Arg Ala Thr Lys Lys Thr Gly Lys Trp Asp Ser Lys Thr 2 2RT Borrelia burgdorferi sensu lato 2Ile Ala Lys Ser Gly Glu Val Thr Val Ala Leu Asn Asp Thr Asn Thr Gln Ala Thr Lys Lys Thr Gly Ala Trp Asp Ser Lys Thr 2 2RT Borrelia burgdorferi sensu lato 2Ile Ser Asn Ser Gly Glu Ile Thr Val Glu Leu Asn Asp Ser Asp Thr Gln Ala Thr Lys Lys Thr Gly Thr Trp Asp Ser Lys Thr 2 2RT Borrelia burgdorferi sensu lato 2Ile Ser Lys Ser Gly Glu Val Thr Ala Glu Leu Asn Asp Thr Asp Thr Gln Ala Thr Lys Lys Thr Gly Lys Trp Asp Ala Gly Thr 2 2RT Borrelia burgdorferi sensu lato 2Asn Gly Thr Lys Leu Glu Gly Ser Ala Val Glu Ile Thr Lys Leu Glu Ile Lys Asn 2orrelia burgdorferi sensu lato 2Ala Gly Thr Asn Leu Glu Gly Asn Ala Val Glu Ile Lys Thr Leu Glu Leu Lys Asn 2orrelia burgdorferi sensu lato 2Ala Gly Thr Asn Leu Glu Gly Lys Ala Val Glu Ile Thr Thr Leu Glu Leu Lys Asn 2orrelia burgdorferi sensu lato 2Ala Gly Thr Asn Leu Glu Gly Thr Ala Val Glu Ile Lys Thr Leu Glu Leu Lys Asn 2orrelia burgdorferi sensu lato 2Leu Asn Asp Thr Asp Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Asn Ser Gly Thr 2orrelia burgdorferi sensu lato 2Leu Asp Asp Ser Asp Thr Thr Gln Ala Thr Lys Lys Thr Gly Lys Asp Ser Lys Thr 2orrelia burgdorferi sensu lato 2Leu Asn Asp Ser Asp Thr Ser Ala Ala Thr Lys Lys Thr Ala Ala Asn Ser Gly Thr 2orrelia burgdorferi sensu lato 2Leu Asn Asp Thr Asp Ser Ser Ala Ala Thr Lys Lys Thr Gly Lys Asn Ser Gly Thr 2orrelia burgdorferi sensu lato 2Leu Asn Asp Thr Asp Ser Ser Ala Ala Thr Lys Lys Thr Ala Ala Asp Ser Lys Thr 2orrelia burgdorferi sensu lato 2Leu Asn Asp Ser Asp Thr Ser Ala Ala Thr Lys Lys Thr Gly Lys Asn Ser Gly Thr 2orrelia burgdorferi sensu lato 2Leu Asn Asp Ser Asp Thr Ser Ala Ala Thr Lys Lys Thr Ala Ala Asp Ser Lys Thr 2BR>* * * * *

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8.