MUTANT FRAGMENTS OF OSPA AND METHODS AND USES RELATING THERETO

Abstract

The present invention relates to a polypeptide comprising a mutant fragment of an outer surface protein A (OspA), a nucleic acid coding the same, a pharmaceutical composition (particularly for use as a medicament of in a method of treating or preventing a Borrelia infection) comprising the polypeptide and/or the nucleic acid, a method of treating or preventing a Borrelia infection and a method of immunizing a subject.

Claims

1.-48. (canceled)

49. A polypeptide comprising a mutant fragment of a Borrelia outer surface protein A (OspA), wherein said mutant OspA fragment comprises SEQ ID NO: 216, or a variant thereof, wherein said variant has at least 95% sequence identity to a wild-type serotype 1 OspA fragment defined by amino acid residues 126-273 of SEQ ID NO: 20, and wherein said variant differs from said wild-type serotype 1 OspA fragment at least by the addition of at least one disulfide bond.

50. The polypeptide according to claim 49, wherein said polypeptide comprises a heterodimer selected from the group consisting of Lip-S1D1-S2D1 (SEQ ID NO: 186), Lip-S2D1-S1D1 (SEQ ID NO: 192), Lip-S1D1-S2D4 (SEQ ID NO: 198), and Lip-S2D4-S1D1 (SEQ ID NO: 203).

51. The polypeptide according to claim 49, wherein said polypeptide consists of a heterodimer selected from the group consisting of Lip-S1D1-S2D1 (SEQ ID NO: 186), Lip-S2D1-S1D1 (SEQ ID NO: 192), Lip-S1D1-S2D4 (SEQ ID NO: 198), and Lip-S2D4-S1D1 (SEQ ID NO: 203).

52. A pharmaceutical composition comprising the polypeptide according to claim 49.

53. The pharmaceutical composition of claim 52, further comprising a pharmaceutically acceptable carrier or excipient.

54. The pharmaceutical composition according to claim 53, wherein said excipient is L-methionine and/or aluminium hydroxide.

55. A pharmaceutical composition comprising Lip-S1D1-S2D1 (SEQ ID NO: 186) and Lip-S5D1-S6D1 (SEQ ID NO: 190).

56. The pharmaceutical composition of claim 55, further comprising a pharmaceutically acceptable carrier or excipient.

57. The pharmaceutical composition according to claim 56, wherein said excipient is L-methionine and/or aluminium hydroxide.

58. A vaccine comprising Lip-S1D1-S2D1 (SEQ ID NO: 186) and Lip-S5D1-S6D1 (SEQ ID NO: 190).

59. The vaccine of claim 58, further comprising a pharmaceutically acceptable carrier or excipient.

60. The vaccine according to claim 59, wherein said excipient is L-methionine and/or aluminum hydroxide.

Description

[0346] The present invention is further illustrated by the following Figures, Tables, Examples and the Sequence listing, from which further features, embodiments and advantages may be taken. As such, the specific modifications discussed are not to be construed as limitations on the scope of the invention. It will be apparent to the person skilled in the art that various equivalents, changes, and modifications may be made without departing from the scope of the invention, and it is thus to be understood that such equivalent embodiments are to be included herein.

[0347] In connection with the present invention

[0348] FIG. 1 shows the amino acid alignment of OspA serotypes 1-6 from Borrelia.

[0349] FIG. 2 schematically shows the production of mutant OspA fragment heterodimers according to the current invention.

[0350] FIG. 3 schematically represents the polypeptide components of one possible pharmaceutical composition of the current invention comprising three different mutant OspA heterodimers, a combination vaccine.

[0351] FIG. 4 shows the chemical structure of Pam.sub.3Cys, an example of a fatty acid substituted cysteine, such as would be found at the N-terminus of lipidated polypeptides of the current invention.

[0352] FIG. 5 shows the binding of antibodies from mice immunized with mutant OspA fragment heterodimer polypeptides of the invention to the cell surface of Borrelia of OspA serotypes 1-6.

[0353] Table 1 shows the thermal stability of the folding of mutant serotype 2 OspA fragments with disulfide bond types from D1 to D5 (for nomenclature, see Table A-4) compared to the wild-type serotype 2 OspA fragment without disulfide bonds (D0).

[0354] Table 2 shows the protection of mice from B. afzelii (strain IS1) infection by the Tick Challenge Method following immunization with mutant serotype 2 OspA fragments with disulfide bond types D1 to D5 (for nomenclature, see Table A-4), including control groups of mice immunized with PBS, full-length OspA or the wild-type serotype 2 OspA fragment (S2D0-His).

[0355] Table 3 shows the protection of mice from B. afzelii (strain IS1) infection by the Tick Challenge Method following immunization with lipidated mutant serotype 2 OspA fragments with disulfide bond types D1, D3 and D4 (Lip-S2D1-His, Lip-S2D3-His and Lip-S2D4-His), including control groups of mice immunized with PBS or full-length OspA protein.

[0356] Table 4 shows the protective capacity of mutant OspA heterodimers of the invention in in vivo Borrelia challenge models. Mice were immunized with Lip-S1D1-S2D1-His, Lip-S4D1-S3D1-His, Lip-S4D1-S3D1 or Lip-S5D1-S6D1-His and challenged with the indicated Borrelia OspA serotype via Tick or Needle Challenge Method, as indicated. The control group in each experiment was immunized with Al(OH).sub.3 adjuvant alone.

[0357] Table 5 shows the protective capacity of the combination vaccine of the invention against challenge in vivo with OspA serotype 1 Borrelia (strain N40 in needle challenge method) and OspA serotype 2 Borrelia (strain IS1 in the tick challenge method). Mice were immunized with the three antigens Lip-S1D1-S2D1, Lip-S4D1-S3D1 and Lip-S5D1-S6D1 together in a 1:1:1 ratio (combination vaccine) or with the indicated control antigens and challenged with Borrelia via Tick or Needle Challenge Method, as indicated. The control group in each experiment was immunized with Al(OH)3 adjuvant alone.

[0358] The figures and tables which may be referred to in the specification are described below in more detail.

[0359] FIG. 1 Amino acid sequence alignment of OspA serotypes one through six (ST1-ST6), excluding the N-terminal lipidation signal sequence (amino acids 1-16), which is truncated during processing. The alignment illustrates that the membrane-associated N-terminal portion of the protein has a more highly-conserved amino acid sequence than the more exposed C-terminal portion. The full-length OspA sequences (excluding the N-terminal lipidation signal sequences, amino acids 1-16, which are not shown in the figure) are as follows: OspA ST3, SEQ ID NO: 21; OspA ST1, SEQ ID NO: 20; OspA ST4, SEQ ID NO: 22; OspA ST5, SEQ ID NO: 23; OspA ST6, SEQ ID NO: 24; OspA ST2, SEQ ID NO: 19.

[0360] FIG. 2 Production of a mutant OspA heterodimer of the invention comprising mutant OspA C-terminal fragments from two different OspA serotypes of Borrelia sp. (A) Schematic representation of a nucleic acid encoding a lipidated mutant OspA heterodimer. The components, from 5 to 3, comprise the coding sequences for a lipidation signal sequence (Lip signal), a small cysteine-containing peptide for N-terminal lipidation (Lipidation peptide=LP), a mutant C-terminal fragment of OspA with two non-native cysteines, a short linker peptide (LN1), followed by a second mutant OspA C-terminal fragment with two non-native cysteines. (B) The intermediate mutant OspA heterodimer polypeptide comprises the nascent product directly following translation of the nucleic acid construct. From the N- to the C-terminus, this polypeptide consists of a lipidation signal sequence (Lip signal), a cysteine-containing peptide for lipidation (LP), a mutant OspA fragment with a non-native disulfide bond, a short linker peptide (LN1), followed by a second mutant OspA fragment with a non-native disulfide bond. (C) The final lipidated mutant OspA heterodimer polypeptide after post-translational modification. The heterodimer, from the N- to the C-terminus, consists of a short cysteine-containing peptide with the N-terminal cysteine lipidated (indicated by Lip), a mutant OspA fragment stabilized by a disulfide bond, a linker peptide (LN1), and a second mutant OspA fragment stabilized by a disulfide bond. The lipidation signal sequence is cleaved off during post-translational modification of the polypeptide as shown.

[0361] FIG. 3 An example of a preferred pharmaceutical composition according to the current invention. Three mutant OspA heterodimers, each comprising mutated OspA fragments from two different Borrelia OspA serotypes are present in the composition, together providing OspA antigens from six different Borrelia OspA serotypes. Such a pharmaceutical composition enables simultaneous immunization against six Borrelia serotypes.

[0362] FIG. 4 Illustration of the chemical structure of Pam.sub.3Cys, an example of a fatty acid substitution of the N-terminal cysteine of full-length wild-type OspA protein as well as of lipidated mutant OspA fragment monomers and heterodimers of the invention. During post-translational modification of a full-length OspA protein or polypeptides of the invention, the N-terminal lipidation signal sequence is cleaved off and fatty acids, most commonly three palmitoyl moieties (Pam.sub.3), are enzymatically covalently attached to the N-terminal cysteine residue (the sulfur atom, S, is indicated by an arrow). The remaining residues of the polypeptide chain, which are located C-terminally from the Pam.sub.3Cys residue, are represented by Xn. (Modified from Bouchon, et al. (1997) Analytical Biochemistry 246: 52-61.)

[0363] FIG. 5 Binding of antibodies from immunized mice to the cell surface of Borrelia spirochetes. Mice were immunized three times with 1 g each of the indicated antigens: Lipidated and His-tagged full-length OspA proteins of OspA serotypes 1-6; Lip-S1D1-S2D1, Lip-S4D1-S3D1 or Lip-S5D1-S6D1 alone, or Lip-S1D1-S2D1, Lip-S4D1-S3D1 and Lip-S5D1-S6D1 together in a 1:1:1 ratio (combination vaccine) at two week intervals and sera were collected at one week after the last dose. Several dilutions of the sera were tested for binding to the cell surface of Borrelia via cell staining and flow cytometry. Fluorescent intensity values observed when staining with sera collected from control mice immunized with Al(OH).sub.3 adjuvant alone were subtracted to account for non-specific binding. (Borrelia used were: B. burgdorferi, OspA serotype 1, strain N40; B. afzelii, OspA serotype 2, strain C; B. garinii, OspA serotype 3, strain D; B. havariensis, OspA serotype 4, strain Fin; B. garinii, OspA serotype 5, strain E; B. garinii, OspA serotype 6, strain B.)

TABLE-US-00007 TABLE 1 Thermal stability of non-lipidated, His-tagged B. afzelii K78 mutant serotype 2 OspA fragments with different placement of disulfide bonds. Mutant serotype 2 OspA fragments with different cysteine bond types (see Table A-4) were solubilized in 50 mM Tris-HCl, 150 mM NaCl (pH 8.0) and tested for thermal stability compared with the wild-type serotype 2 OspA fragment (S2D0). The presence of a disulfide bond resulted in an increased melting temperature compared to the wild-type serotype 2 OspA fragment Serotype 2 OspA Melting mutant fragment SEQ ID NO: temperature ( C.) S2D0-His* 1 47.6 S2D1-His 2 70.4 S2D2-His 3 54.6 S2D3-His 4 58.6 S2D4-His 5 58.4 S2D5-His 6 53.8 *see Tables A-4 and A-5 for nomenclature.

TABLE-US-00008 TABLE 2 Protective capacity of decreasing doses of non-lipidated His-tagged mutant serotype 2 OspA fragments against B. afzelii (serotype 2) infection by the Tick Challenge Method. Five non-lipidated His-tagged mutant serotype 2 OspA fragments were tested for protective capacity at two different doses (30 g and 5 g) and compared with the wild-type serotype 2 OspA fragment. Groups of mice immunized with Al(OH).sub.3 adjuvant alone or with non-lipidated full-length serotype 2 OspA served as negative and positive controls, respectively. All antigens were His-tagged and non-lipidated. The data presented combine the results of several experiments performed under identical conditions. 3 x 30 g 3 x 5 g Tick challenge (11 exper- (4 exper- (OspA serotype 2: iments) iments) B.afzelii, strain Infected/ Infected/ Immunogen IS1) total total Al(OH).sub.3 adjuvant alone Tick (OspA-ST2) 58/62 20/23 Full-length OspA K78-His Tick (OspA-ST2) 1/72 1/25 (SEQ ID NO: 209) S2D0-His Tick (OspA-ST2) 15/20 8/16 (SEQ ID NO: 1) S2D1-His Tick (OspA-ST2) 1/26 1/25 (SEQ ID NO: 2) S2D2-His Tick (OspA-ST2) 0/26 4/26 (SEQ ID NO: 3) S2D3-His Tick (OspA-ST2) 0/34 1/21 (SEQ ID NO: 4) S2D4-His Tick (OspA-ST2) 2/30 4/27 (SEQ ID NO: 5) S2D5-His Tick (OspA-ST2) 5/35 2/11 (SEQ ID NO: 6)

TABLE-US-00009 TABLE 3 Protective capacity of decreasing doses of lipidated His-tagged mutant serotype 2 OspA fragments against B. afzelii infection by the Tick Challenge Method. Three lipidated His-tagged mutant serotype 2 OspA fragments with different disulfide bond types were tested for protective capacity at three different doses (3.0 g, 1.0 g and 0.3 g). Groups of mice immunized with Al(OH).sub.3 adjuvant alone or with non-lipidated full-length serotype 2 OspA served as negative and positive controls, respectively. The data presented combine the results of several experiments performed under identical conditions. Tick challenge 3 x 3.0 g 3 x 1.0 g 3 x 0.3 g (OspA serotype 2: (5 experiments) (5 experiments) (4 experiments) Immunogen B.afzelii, strain IS1) Infected/total Infected/total Infected/total Al(OH).sub.3 adjuvant alone Tick (OspA-ST2) 58/59 (control for all doses) Full-length OspA K78-His Tick (OspA-ST2) 0/14 0/21 1/20 (SEQ ID NO: 209) Lip-S2D1-His Tick (OspA-5T2) 0/17 5/31 1/29 (SEQ ID NO: 141) Lip-S2D3-His Tick (OspA-ST2) 1/15 1/12 5/19 (SEQ ID NO: 143) Lip-S2D4-His Tick (OspA-ST2) 0/8 0/25 0/34 (SEQ ID NO: 144)

TABLE-US-00010 TABLE 4 Protective capacity of mutant OspA heterodimers of the invention against in vivo Borrelia challenge via Needle or Tick Challenge Methods. Groups of mice were immunized three times at two week intervals with the indicated doses of OspA heterodimer or Al(OH).sub.3 adjuvant alone. Immunogens used were Lip-S1D1-S2D1-His (challenged with Borrelia OspA-ST1, Experiments 1-3), Lip-S1D1-S2D1-His, Lip-S4D1-S3D1-His and Lip-S5D1-S6D1-His, separately (challenged with Borrelia OspA-ST2, Experiments 4-6), Lip-S4D1-S3D1 (challenged with Borrelia OspA-ST4, Experiments 7 and 8) and Lip-S5D1-S6D1-His (challenged with Borrelia OspA-ST5, Experiments 9 and 10; challenged with Borrelia OspA-ST6, Experiments 11 and 12). Immunized mice were challenged two weeks after the last immunization via Tick or Needle Challenge Models as indicated. Needle challenge (OspA-serotype 1: B. Infected/Total Immunogen Dose burgdorferi s.s., strain N40) Exp. 1 Exp. 2 Exp. 3 Lip-S1D1-S2D1-His 3 5.0 g Needle (OspA-ST1) 0/10*** 0/9*** 4/10** (SEQ ID NO: 49) Al(OH).sub.3 adjuvant alone Needle (OspA-ST1) 10/10 8/10 10/10 Tick challenge (OspA-serotype Immunogen Dose 2: B.afzelii, strain IS1) Exp. 4 Exp. 5 Exp. 6 Lip-S1D1-S2D1-His 3 2.0 g Tick (OspA-ST2) 0/10*** 0/9*** 0/6*** (SEQ ID NO: 49) Lip-S4D1-S3D1-His 3 2.0 g Tick (OspA-ST2) 0/9*** 2/7* 0/6*** (SEQ ID NO: 81) Lip-S5D1-S6D1-His 3 2.0 g Tick (OspA-ST2) 0/7*** 0/9*** 0/6*** (SEQ ID NO: 65) Al(OH).sub.3 adjuvant alone Tick (OspA-ST2) 9/9 8/8 7/7 Needle challenge (OspA-serotype 4: B. Immunogen Dose bavariensis, strain Scf) Exp. 7 Exp. 8 Lip-S4D1-S3D1 3 5.0 g Needle (OspA-ST4) 2/10** 1/10*** (Seq ID No: 194) Al(OH).sub.3 adjuvant alone Needle (OspA-ST4) 9/10 9/10 Needle challenge Immunogen Dose (OspA-serotype 5: B. garinii) Exp. 9 Exp. 10 Lip-S5D1-S6D1-His 3 5.0 g Needle (OspA-ST5) 1/10 2/10 (SEQ ID NO: 65) Al(OH).sub.3 adjuvant alone Needle (ST5) 6/10 6/10 Needle challenge Immunogen Dose (OspA-serotype 6: B. garinii) Exp. 11 Exp. 12 Lip-S5D1-56D1-His 3 5.0 g Needle (OspA-ST6) 2/10** 2/10*** (SEQ ID NO: 65) Al(OH).sub.3 adjuvant alone Needle (OspA-ST6) 9/10 10/10 P-value; Fisher's exact test, two tailed. *significant (<0.05), **highly significant (<0.01), ***extremely significant (<0.001)

TABLE-US-00011 TABLE 5 Protective capacity of the mutant OspA heterodimer combination vaccine of the invention against OspA serotype 1 and serotype 2 Borrelia challenge. Groups of mice were immunized three times with the indicated doses of immunogen or Al(OH).sub.3 adjuvant alone at two- week intervals. Immunogens used were a 1:1:1 combination of the mutant OspA heterodimers Lip-S1D1-S2D1, Lip-S4D1-S3D1 and Lip-S5D1-S6D1 (combination vaccine), Lip-S1D1-S2D1, Lip-OspAl-His and Chimeric OspA ST1/ST2. Immunized mice were challenged two weeks after the last immunization via the Tick Challgenge Method (ST2, Experiments 13 and 14) or the Needle Challenge Method (ST1, Experiments 15 and 16). Tick challenge (OspA-serotype 2: B Infected/Total Immunogen Dose afzelii, strain IS1) Exp. 13 Exp. 14 Lip-S1D1-S2D1 3 5.0 g Tick (OspA-ST2) 0/6*** 0/7** (SEQ ID NO: 186) Combination vaccine: Lip-S1D1-S2D1 3 5.0 g Tick (OspA-ST2) 0/9*** 0/6** (Seq ID No: 186) Lip-S4D1-S3D1 3 5.0 g (Seq ID No: 194) Lip-S5D1-S6D1 3 5.0 g (Seq ID No: 190) Al(OH).sub.3 adjuvant alone Tick (OspA-ST2) 7/7 6/7 Needle challenge (OspA-serotype 1: B. burgdoileri s.s., Immunogen Dose strain ZS7) Exp. 15 Exp. 16 Lip-SiD1-S2D1 3 1.0 g Needle (OspA-ST1) 0/10*** 0/10*** (Seq ID No: 186) Lip-OspAl-His 3 1.0 g Needle (OspA-ST1) 0/10*** 0/10*** (Seq ID No: 210) Chimeric OspA ST1/ST2 3 1.0 g Needle (OspA-ST1) 0/10*** 0/10*** (Seq ID No: 212) Combination vaccine: Lip-S1D1-S2D1 (Seq ID No: 186) 3 1.0 g Needle (OspA-STI) 0/10*** 0/10*** Lip-S4D1-S3D1 (Seq ID No: 194) 3 1.0 g Lip-S5D1-S6D1 (Seq ID No: 190) 3 1.0 g Al(OH).sub.3 adjuvant alone Needle (OspA-ST1) 10/10 10/10 P-value; Fisher's exact test, two tailed. *significant (<0.05), **highly significant (<0.01), ***extremely significant (<0.001)

EXAMPLES

Example 1. Assessment of Thermal Stability of Mutant Serotype 2 OspA Fragments

[0364] Experimental Procedures

[0365] Thermal Stability

[0366] The melting temperatures (T.sub.m) of non-lipidated mutant serotype 2 OspA fragment monomers were determined by the fluorescence-based thermal shift assay described by Pantoliano, et al. (J. Biomol Screen 6:429-440 (2001)). The fluorescent dye SYPRO Orange protein gel stain (supplied as a 5000 concentrate in DMSO by Sigma, U.S.A.) was used to monitor protein unfolding. In each well, 7.5 L of SYPRO Orange (diluted 1:1000 from the stock solution) and 17.5 L of a solution of protein (1 g or 2 g) in buffer were combined. The protein samples were heated from 25 C. to 95 C. at a rate of 0.2 C./10 sec in the CFX96 Real-time Detection System (Bio-Rad, USA) and fluorescent changes were monitored. Fluorescence intensity was measured with excitation and emission wavelengths of 490 and 575 nm, respectively. The T.sub.m was determined using the Bio-Rad CFX Manager 2.0 program. The T.sub.in values of non-lipidated His-tagged serotype 2 OspA mutant fragments were measured in four different buffer systems: 50 mM Tris-HCl, 150 mM NaCl (pH 9.0); 50 mM Tris-HCl, 150 mM NaCl (pH 8.0); PBS (pH 7.4); and 25 mM HEPES, 150 mM NaCl (pH 6.5), using the non-lipidated serotype 2 OspA wild-type fragment (S2D0) as a control.

[0367] Results

[0368] In all cases, mutant serotype 2 OspA fragments with an introduced cysteine bond had higher T.sub.m than the wild-type serotype 2 OspA fragment (S2D0) (see Table 1). The T.sub.m was tested in four different buffer systems with similar results (data for proteins dissolved in 50 mM Tris-HCl, 150 mM NaCl (pH 8.0) is shown in Table 1), indicating that the stability of the proteins is similar over a wide pH range. This result lends credence to the hypothesis that the introduced disulfide bond stabilizes the OspA fragment.

Example 2. Assessment of the Protective Capacity of Non-Lipidated His-Tagged Mutant Serotype 2 OspA Fragment Monomers in the Tick Challenge Method (ST2, B. afzelii)

[0369] Experimental Procedures

[0370] Cloning and Expression of Recombinant Proteins

[0371] The wild-type serotype 2 OspA fragment as well as the mutant serotype 2 OspA fragments with cysteine bond types 1-5 (SEQ ID NOs: 1, 2, 3, 4, 5 and 6, respectively), were codon-optimized for E. coli expression by GenScript, USA. The non-lipidated mutant serotype 2 OspA fragments were C-terminally His-tagged for purification purposes. Gene fragments were cloned into the pET28b(+) vector (Novagen, USA), a vector containing a Kanamycin resistance cassette as well as a T7 promoter. The monomers were expressed in BL21 Star (DE3) cells (Invitrogen, USA) at 37 C. by the addition of IPTG. Cells were collected after 4 h by centrifugation and the pellet was stored at 70 C. for up to 12 months prior to further processing.

[0372] Purification of Non-Lipidated His-Tagged Wild-Type and Mutant OspA Fragment Monomer Proteins

[0373] Cells were disrupted mechanically by high-pressure homogenization and the soluble fraction containing the His-tagged OspA fragments was applied to a Ni-sepharose column (Ni Sepharose 6 Fast Flow; GE Healthcare, United Kingdom) and the His-tagged OspA fragments were eluted on an Imidazole gradient (0-250 mM). Pooled fractions were further purified over a gel filtration column (Superdex 200, GE Healthcare) followed by a buffer exchange column (Sephadex G-25, GE Healthcare). His-tagged OspA fragment peaks were pooled on the basis of the analytical size exclusion column and reversed phase chromatography. After sterile filtration, the purified proteins were stored at 20 C. until formulation.

[0374] Immunization of Mice

[0375] Female C3H/HeN (H-2.sup.k) mice were used for all studies (Harlan, Italy). Prior to each challenge, groups of five 8-week-old mice were bled via the tail vein and pre-immune sera were prepared and pooled. Five non-lipidated mutant serotype 2 OspA fragment proteins (S2D1-5, SEQ ID NOs: 2, 3, 4, 5 and 6, respectively), were tested in fifteen separate experiments. Three subcutaneous (s.c.) immunizations of 100 L, were administered at two week intervals. Doses used were 30 and 5 g of the respective protein, tested in 11 and 4 experiments respectively. All formulations included aluminium hydroxide (Al(OH).sub.3) at a final concentration of 0.15%. One week after the third immunization, blood was collected and hyper-immune sera were prepared. In each experiment, one group injected with PBS formulated with Al(OH).sub.3 was included as a negative control and one group of mice was immunized with S2D0, the wild-type C-terminal OspA fragment from B. afzelii strain K78 (SEQ ID NO: 1). Another group immunized with a non-lipidated full-length wild-type OspA protein from B. afzelii, strain K78 (SEQ ID NO: 209), also formulated with 0.15% Al(OH).sub.3, was included as positive control in each animal study. All animal experiments were conducted in accordance with Austrian law (BGB1 Nr. 501/1989) and approved by Magistratsabteilung 58.

[0376] Tick Challenge of Immunized Mice and Collection of Sera and Tissues (Herein Referred to Also as Tick Challenge Method)

[0377] Tick challenge of immunized mice was done two weeks after the last immunization. In order to challenge the immunized mice with B. afzelii, the hair on the back of each mouse was removed with Veet Cream (Reckitt Benckiser, United Kingdom) and a small ventilated container was glued to the skin with super glue (Pattex, Germany). Thereafter, one or two L ricinus nymphs infected with B. afzelii, strain IS1, were applied per mouse, allowed to attach and feed to depletion. The feeding status was monitored daily for each individual tick and only mice where at least one fully-fed tick was collected were included in the final readout. No distinction was made between mice where one or two fully-fed ticks were collected.

[0378] Six weeks after the tick application, blood was collected by orbital bleeding and final sera were prepared and used for VlsE ELISA analysis to determine infection status. The mice were then sacrificed by cervical dislocation and one ear from each mouse was collected, DNA extracted and subjected to nested PCR analysis to identify Borrelia in tissue.

[0379] Infection Readout

[0380] Only mice where the applied tick(s) fed to completion and could be collected were included in the final readout of the experiment. The mice were sacrificed 6 weeks after tick application and organs as well as final sera were collected. The final infection readout was based on two different analyses (nested PCR targeting the 16S-23S intergenic spacer and VlsE (IR6) ELISA as described in detail below).

[0381] Nested PCR Targeting the 16S-23S Intergenic Spacer

[0382] One ear from each mouse was subjected to DNA extraction and purification using the DNeasy Blood and Tissue Kit (Qiagen, Germany) according to the manufacturer's instructions, with the following modification. Each ear was digested over night at 60 C. in recombinant Proteinase K, PCR grade (Roche, 14-22 mg/mL). The DNA was eluted in 50 L deionized sterile water and stored at 20 C. until further analysis. As a negative control, one empty purification column was included in each DNA extraction and purification and the eluate subjected to nested PCR. All DNA extracts were screened for the presence of Borrelia DNA by a nested PCR procedure, comprising 40 cycles of 94 C. for 30 s, 56 C. for 30 s and 72 C. for 60 s using the primers; Forward 5-GTATGTTTAGTGAGGGGGGTG-3 (SEQ ID NO: 26) and Reverse 5-GGATCATAGCTCAGGTGGTTAG-3 (SEQ ID NO: 27). From the reaction volume of 10 L, 1 L was used as template for the nested PCR reaction. The nested PCR step comprised 25 cycles of 94 C. for 30 s, 60 C. for 30 s and 72 C. for 60 s using the primers; Forward nested 5-AGGGGGGTGAAGTCGTAACAAG-3 (SEQ ID NO: 28) and Reversed nested 5-GTCTGATAAACCTGAGGTCGGA-3 (SEQ ID NO: 29). Of the final reaction volume, 5 L was separated on a 1% agarose gel containing ethidium bromide and bands were visualized in UV-light.

[0383] In each PCR analysis, DNA purified from an in vitro grown culture of B. afzelii strain K78 was used as a positive control template. In addition, PBS was used instead of extracted DNA as negative control. Five microliters of the final product was separated on a 1% agarose gel containing ethidium bromide and bands were visualized in UV-light.

[0384] ELISA with the Invariable Region 6 (IR6) of the Variable Major Protein-Like Sequence E Protein (VlSE)

[0385] A biotinylated 25-mer peptide (MKKDDQIAAAMVLRGMAKDGQFALK) (SEQ ID NO: 30) derived from the sequence of B. garinii strain IP90 was used for analysis (Liang F T, et al. (1999) J Immunol 163:5566-73). Streptavidin pre-coated 96-well ELISA plates (Nunc, Denmark) were coated with 100 L/well (1 g/mL) biotinylated peptide in PBS supplemented with 0.1% Tween 20 (PBS/0.1T). The plates were incubated overnight at 4 C. After coating with the peptide, the plates were washed once with PBS/0.1T. The plates were then blocked for one hour at room temperature (RT) with 100 L/well of PBS+2% BSA, before being washed again with PBS/0.1T. Reactivity of post-challenge sera to the peptide was tested at 1:200, 1:400 and 1:800 dilutions in PBS+1% BSA. Plates were incubated for 90 min at RT before being washed three times with PBS/0.1T. Each well then received 50 L of 1.3 g/mL polyclonal rabbit anti-mouse IgG conjugated to HRP (Dako, Denmark) in PBS+1% BSA. The plates were then incubated for 1 h at RT. After three washes with PBS/0.1T, ABTS (50 L/well) was added as substrate (Sigma-Aldrich, USA) and color was allowed to develop for 30 min. Absorbance was measured at 405 nm. All sera were tested in duplicate; negative controls included PBS instead of sera, as well as plates not coated with the peptide. Sera from mice shown to be culture positive for B. afzelii infection were used as positive controls.

[0386] Results

[0387] Levels of Protection in the Tick Challenge Method

[0388] High levels of protection were observed for all five stabilized OspA B. afzelii fragments at both of the doses tested (30 g and 5 g, see Table 2). The high infection rates in the PBS control group indicate that the ticks were infected with high frequency. Additionally, the positive control, non-lipidated full-length OspA from B. afzelii strain K78, was very protective. Together these control groups indicate the high reliability of the experimental readout.

[0389] Protection results from experiments testing 30 g doses (11 total experiments) and 5 g doses (4 total experiments) are summarized in Table 2. The two methods employed to verify infection, namely VlsE ELISA and nested PCR, gave virtually identical results (data not shown), demonstrating the robustness of these readout methods for assessing infection in the Tick Challenge Method.

Example 3. Assessment of the Protective Capacity of Lipidated His-Tagged Mutant Serotype 2 OspA Fragment Monomers Against In Vivo Borrelia Challenge Via the Tick Challenge Method (ST2, B. afzelii)

[0390] Experimental Procedures

[0391] Cloning and Expression of Lipidated His-Tagged Mutant OspA Fragment Proteins

[0392] The serotype 2 mutant OspA fragments with cysteine bond types 1, 3 and 4 (SEQ ID NOs: 141, 143 and 144, respectively) were modified by the addition of a lipidation signal sequence derived from OspA (SEQ ID NO: 14) and followed directly C-terminally by a CKQN peptide (SEQ ID NO: 211) to provide an N-terminal cysteine for lipidation. All mutant OspA fragments were C-terminally histidine-tagged for purification purposes. Gene fragments were cloned into the pET28b(+) vector (Novagen), a vector containing a Kanamycin resistance cassette as well as a T7 promoter. The lipidated monomers were expressed in BL21 Star (DE3) cells (Invitrogen) and after induction by IPTG, the growth temperature of the cells was lowered from 37 C. to 25 C. to promote efficient post-translational processing of the proteins. Cells were collected after 4 h by centrifugation and the pellet was stored at 70 C. for up to 12 months prior to further processing.

[0393] Purification of Lipidated His-Tagged Wild-Type and Mutant OspA Fragment Monomer Proteins

[0394] Cells were disrupted mechanically by high-pressure homogenization and the lipidated His-tagged OspA fragment monomer polypeptides were enriched in the lipid phase by phase separation, using Triton X-114 as detergent. Subsequently, the diluted detergent phase (20 to 30 fold) was applied to a Ni-sepharose column (Ni Sepharose 6 Fast Flow; GE Healthcare) and the lipidated His-tagged OspA fragments were eluted by Imidazole gradient (0-250 mM) elution. Pooled fractions were further purified over a gel filtration column (Superdex 200, GE Healthcare) followed by a buffer exchange column (Sephadex G-25, GE Healthcare). Lipidated His-tagged OspA fragment peaks were pooled on the basis of the analytical size exclusion column and reversed phase chromatography. After sterile filtration, the purified proteins were stored at 20 C. until formulation.

[0395] Immunization of Mice

[0396] Three lipidated mutant OspA proteins (Lip-S2D1-His, Lip-S2D3-His and Lip-S2D4-His) were expressed and purified as described above. In vivo protection studies were performed as described in Example 2 using Al(OH).sub.3-adjuvant alone and non-lipidated full-length serotype 2 OspA as negative and positive controls, respectively. All immunogens were formulated with 0.15% Al(OH).sub.3. Mice were injected subcutaneously three times at two week intervals with formulations containing 3.0 g, 1.0 g or 0.3 g antigen and challenged with B. afzelii-infected ticks (strain IS1) two weeks after the last immunization. Mice were sacrificed six weeks following tick challenge and infection was assessed.

[0397] Results

[0398] Levels of Protection in the Tick Challenge Method

[0399] All three lipidated mutant OspA fragments conferred very high levels of protection from B. afzelii challenge even at the lowest tested dose (Table 3). Infection rates in the Al(OH).sub.3-adjuvant alone immunized mice were high, indicating that the ticks were infected to a high frequency. The positive control antigen, full-length non-lipidated OspA from B. afzelii strain K78, was also very protective. Together, these control groups indicate the high reliability of the method of infection and thus give high credibility to the results observed following immunization with the lipidated mutant OspA fragments.

Example 4. Assessment of the Protective Capacity of Mutant OspA Heterodimers of the Invention Against In Vivo Borrelia Challenge Via the Needle or Tick Challenge Methods

[0400] Experimental Procedures

[0401] Cloning and Expression of Lipidated His-Tagged Mutant OspA Fragment Heterodimers

[0402] The mutant OspA fragment monomers from B. burgdorferi s.s. strain B31, B. afzelii strain K78, B. garinii strain PBr, B. bavariensis strain PBi, B. garinii strain PHEi and B. garinii strain DK29 were codon-optimized for E. coli expression by GenScript, USA. The hLFA-1-like epitope (aa 164-174, SEQ ID NO: 17) of the OspA from B. burgdorferi s.s. strain B31 was replaced by a non-hLFA-1-like sequence NFTLEGKVAND from B. afzelii strain K78 (SEQ ID NO: 18). The lipidation signal sequence added to the mutant OspA fragment heterodimers was derived from the E. coli major outer membrane lipoprotein, Lpp, and was followed directly C-terminally by a CSS peptide to provide an N-terminal cysteine for lipidation. The mutant OspA fragment heterodimers were generated by fusing different mutant OspA fragment monomers as described above via a 21 amino acid linker sequence, originating from two separate loop regions of the N-terminal half of OspA from B. burgdorferi s.s. strain B31 (LN1; aa 65-74 and aa 42-53 with an amino acid exchange of D53S, SEQ ID NO: 184). The heterodimers were constructed with a His-tag for purification purposes. Gene fragments were cloned into the pET28b(+) vector (Novagen), a vector containing a Kanamycin resistance cassette as well as a T7 promoter. The lipoproteins of the stabilized heterodimers were expressed in BL21 Star (DE3) cells (Invitrogen) and after induction by IPTG, the growth temperature of the cells was lowered from 37 C. to 25 C. to promote efficient post-translational processing of the proteins. Cells were collected after 4 h by centrifugation and the pellet was stored at 70 C. for up to 12 months prior to further processing.

[0403] Purification of Lipidated His-Tagged Mutant OspA Fragment Heterodimers

[0404] Cells were disrupted mechanically by high-pressure homogenization and the lipidated His-tagged mutant OspA fragment heterodimers were enriched in the lipid phase by phase separation, using Triton X-114 as detergent. Subsequently, the diluted detergent phase (20 to 30 fold) was applied to a Ni-sepharose column (Ni Sepharose 6 Fast Flow; GE Healthcare) and the lipidated His-tagged OspA heterodimers were eluted by Imidazole gradient (0-250 mM) elution. Pooled fractions were further purified over a gel filtration column (Superdex 200, GE Healthcare) followed by a buffer exchange column (Sephadex G-25, GE Healthcare). The lipidated His-tagged mutant OspA heterodimer peaks were pooled on the basis of the analytical size exclusion column and reversed phase chromatography. After sterile filtration, the purified heterodimers were stored at 20 C. until formulation.

[0405] Cloning and Expression of Lipidated Non-His-Tagged Mutant OspA Fragment Heterodimers

[0406] The constructs made as described as above were used for the generation of constructs without a His-tag by the introduction of a stop codon by PCR amplification. Gene fragments were cloned into the pET28b(+) vector (Merck Millipore), a vector containing a kanamycin resistance cassette as well as a T7 promoter. The lipoproteins of the stabilized heterodimers were expressed in BL21 Star (DE3) cells (Invitrogen) and after induction by IPTG, the growth temperature of the cells was lowered from 37 C. to 25 C. to promote efficient post-translational processing of the proteins. Cells were collected after 4 h by centrifugation and the pellet was stored at 70 C. for up to 12 months prior to further processing.

[0407] Purification of Lipidated Non-His-Tagged Mutant OspA Fragment Heterodimers

[0408] Cells were disrupted mechanically by high-pressure homogenization and the lipidated mutant OspA fragment heterodimers were enriched in the lipid phase by phase separation, using Triton X-114 as detergent. Subsequently, the diluted detergent phase was subjected to anion exchange chromatography operated in non-binding mode. The resulting flow-through was loaded on a hydroxyapatite column (Bio-Rad) and the lipidated proteins were eluted from the column by a linear salt gradient. The eluate was subjected to further purification over a DEAE-Sepharose column (GE Healthcare) in non-binding mode followed by gel filtration column (Superdex 200, GE Healthcare) for buffer exchange. The lipidated mutant OspA heterodimer peaks were pooled on the basis of the analytical size exclusion column and SDS-PAGE. After sterile filtration, the purified heterodimers were stored at 20 C. until formulation.

[0409] Immunization of Mice

[0410] Female C3H/HeN mice (Janvier, France) were used for all studies. Prior to each challenge, groups of ten 8-week-old mice were bled via the facial vein and pre-immune sera were prepared and pooled. Three subcutaneous (s.c.) immunizations of 100 L each were administered at two week intervals. Each dose contained the amount of immunogen indicated in Table 4 (dose), formulated with aluminium hydroxide (Al(OH).sub.3) at a final concentration of 0.15%. One week after the third immunization, blood was collected from the facial vein and hyper-immune sera were prepared. In each experiment, one group immunized with Al(OH).sub.3 alone was included as a negative control. All animal experiments were conducted in accordance with Austrian law (BGB1 Nr. 501/1989) and approved by Magistratsabteilung 58.

[0411] Tick Challenge of Immunized Mice and Collection of Sera and Tissues (Herein Referred to Also as Tick Challenge Method)

[0412] In order to challenge the immunized mice with B. afzelii, the hair of the back of each mouse was removed with Veet Cream (Reckitt Benckiser) and a small ventilated container was glued to the skin with super glue (Pattex). Thereafter, one or two I. ricinus nymphs infected with B. afzelii, strain IS1, were applied per mouse, allowed to attach and feed until they were fully engorged and dropped off. The feeding status was monitored daily for each individual tick and only mice from which at least one fully-fed tick was collected were included in the final readout.

[0413] Needle Challenge of Immunized Mice with In Vitro Grown Borrelia

[0414] Two weeks after the last immunization, the mice were challenged s.c. with Borrelia diluted in 100 L Borrelia growth medium (BSK II). The challenge doses were strain-dependent, the virulence of the individual strains being assessed by challenge experiments for determination of ID.sub.50. Doses employed for needle challenge experiments ranged from 20 to 50 times the Ip.sub.50.

[0415] Sacrifice of Mice and Collection of Material

[0416] Four weeks after needle challenge with the indicated Borrelia spp. or six weeks after tick challenge with B. afzelii, mice were sacrificed by cervical dislocation. The blood was collected by orbital bleeding and final sera were prepared and used for VlsE ELISA to determine infection status. In addition, one ear from each mouse was collected, and DNA was extracted and subjected to quantitative PCR (qPCR) for identification of Borrelia. The final infection readout was based on two different analyses (VlsE ELISA and qPCR targeting recA).

[0417] ELISA with the Invariable Region 6 (IR6) of VlsE

[0418] A biotinylated 25-mer peptide (MKKDDQIAAAMVLRGMAKDGQFALK) derived from the sequence of B. garinii strain IP90 was used for the analysis (Liang F T, Alvarez A L, Gu Y, Nowling J M, Ramamoorthy R, Philipp M T. An immunodominant conserved region within the variable domain of VlsE, the variable surface antigen of Borrelia burgdorferi. J Immunol 1999; 163:5566-73). Streptavidin pre-coated 96-well ELISA plates (Nunc), were coated with 100 L/well (1 g/mL) peptide in PBS supplemented with 0.1% Tween (PBS/0.1T). The plates were incubated overnight at 4 C. After coating with the peptide, the plates were washed once with PBS/0.1T. The plates were then blocked for one hour at room temperature (RT) with 100 L/well of PBS+2% BSA, before being washed again with PBS/0.1T. Reactivity of post-challenge sera to the peptide was tested at 1:200, 1:400 and 1:800 dilutions in PBS+1% BSA. Plates were incubated for 90 min at RT before being washed three times with PBS/0.1T. Each well then received 50 L of 1.3 g/mL polyclonal rabbit anti-mouse IgG conjugated to HRP (Dako) in PBS+1% BSA. The plates were then incubated for 1 h at RT. After three washes with PBS/0.1T, ABTS (50 L/well) was added as substrate (Sigma-Aldrich) and color was allowed to develop for 30 min. Absorbance was measured at 405 nm. All sera were tested in duplicate. Negative controls included PBS instead of sera as well as plates not coated with the peptide. Sera from mice shown to be culture positive for B. afzelii infection were used as positive controls.

[0419] qPCR Targeting recA

[0420] Oligonucleotide primers were designed for the recA gene in a manner that they could be used in qPCR for identification of all relevant Borrelia species causing Lyme borreliosis (forward: CATGCTCTTGATCCTGTTTA, reverse: CCCATTTCTCCATCTATCTC). The recA fragment was cloned from the B. burgdorferi s.s. strain N40 into pET28b(+), to be used as standard in each reaction. The chromosomal DNA extracted from mouse ears was diluted 1:8 in water in order to reduce matrix effects observed with undiluted DNA. A master mix consisting of 10 L SSoAdvanced SYBR Green Supermix, 0.3 L of each primer (10 M), and 7.4 L water was prepared for each experiment. Eighteen L of master mix was mixed with 2 L of the diluted DNA extracted from either bladder or car in micro-titer plates and the DNA was amplified using a CFX96 real-time PCR detection system (Bio-Rad, USA). The DNA was denatured for 3 minutes at 95 C., followed by 50 cycles of 15 seconds at 95 C. and 30 seconds at 55 C. After amplification, the DNA was prepared for the melting curve analysis by denaturation for 30 seconds at 95 C. followed by 2 minutes at 55 C. The melting curve analysis was performed by 5 seconds incubation at 55 C., with a 0.5 C. increase per cycle, and 5 seconds at 95 C. On each plate, four no-template controls (NTC) were included as well as a standard curve in duplicate with template copy numbers ranging from 10 to 10,000.

[0421] Results

[0422] Lipidated mutant OspA fragment heterodimers were tested for protective capacity in twelve separate experiments. Mice were challenged with either B. burgdorferi s.s., strain N40, OspA serotype 1 (ST1, needle challenge) or B. afzelii strain IS1, OspA serotype 2 (ST2, tick challenge) in three experiments each or B. bavariensis, strain Scf, OspA serotype 4 (ST4, needle challenge), B. garinii, strain A, OspA serotype 5 (ST5, needle challenge) or B. garinii, strain B, OspA serotype 6 (ST6, needle challenge) in two experiments each. In all experiments, a group of mice immunized with Al(OH).sub.3 adjuvant alone served as a negative control group. For challenge with ticks, 1-2 ticks were applied per mouse and only mice from which at least one tick fed until fully engorged were included in the final readout. However, no distinction was made between mice from which one or two fully-fed ticks were collected. The protection data from the twelve experiments are summarized in Table 4. The lipidated His-tagged OspA heterodimer (Lip-S1D1-S2D1-His) showed highly statistically-significant protection (Fisher's exact test, two-tailed) in all six experiments against both OspA serotype 1 and OspA serotype 2 challenge as compared to the negative control group. Surprisingly, immunization with Lip-S4D1-S3D1-His and Lip-S5D1-S6D1-His also conferred a high protective capacity against OspA serotype 2 challenge (Experiments 4-6), indicating that there can be a cross-protective effect of immunization with other serotypes of the mutant OspA fragments. Furthermore, immunization with Lip-S4D1-S3D1 conferred statistically-significant protection against needle challenge with OspA serotype 4 Borrelia (Experiments 7 and 8). Finally, immunization with Lip-S5D1-S6D1-His conferred protection against needle challenge with both OspA serotype 5 (Experiments 9 and 10) and OspA serotype 6 (Experiments 11 and 12). The infectious status of each mouse was determined using VlsE ELISA in combination with recA qPCR. A mouse was regarded as infected when at least one method gave a positive result.

[0423] In conclusion, immunization with mutant OspA fragment heterodimer polypeptides of the invention confers protection against all Borrelia serotypes tested and also may provide cross-protection in some cases.

[0424] The lipidated His-tagged OspA heterodimer (Lip-S1D1-S2D1-His) showed highly statistically-significant protection (Fisher's exact test, two-tailed) in all six experiments against both OspA serotype 1 and OspA serotype 2 challenge as compared to the negative control group. Surprisingly, immunization with Lip-S4D1-S3D1-His and Lip-S5D1-S6D1-His also conferred a high protective capacity against OspA serotype 2 challenge (Experiments 4-6), indicating that there can be a cross-protective effect of immunization with other serotypes of the mutant OspA fragments. Furthermore, immunization with Lip-S4D1-S3D1 conferred statistically-significant protection against needle challenge with OspA serotype 4 Borrelia (Experiments 7 and 8). Finally, immunization with Lip-S5D1-S6D1-His conferred protection against needle challenge with both OspA serotype 5 (Experiments 9 and 10) and OspA serotype 6 (Experiments 11 and 12). The infectious status of each mouse was determined using VlsE ELISA in combination with recA qPCR. A mouse was regarded as infected when at least one method gave a positive result.

[0425] In conclusion, immunization with mutant OspA fragment heterodimer polypeptides of the invention confers protection against all Borrelia serotypes tested and also may provide cross-protection in some cases.

Example 5. Assessment of the Protective Capacity of a 1:1:1 Combination Vaccine of the Mutant OspA Heterodimers of the Invention Against In Vivo OspA Serotype 1 and Serotype 2 Borrelia Challenge Via the Needle Challenge or Tick Challenge Methods

[0426] Experimental Procedures

[0427] Immunization of Mice

[0428] Female C3H/HeN mice (Janvier, France) were used for all studies. Prior to each challenge, groups of ten 8-week-old mice were bled via the facial vein and pre-immune sera were prepared and pooled. Three s.c. immunizations of 100 L each were administered at two week intervals. Groups of mice were immunized with the combination vaccine consisting of 1 g each of Lip-S1D1-S2D1, Lip-S4D1-S3D1 and Lip-S5D1-S6D1. Three other OspA-based antigens were included in the challenge experiments: Lip-OspA1-His (full-length serotype 1 OspA, lipidated and his-tagged), lipidated chimeric OspA ST1/ST2* and Lip-S1D1-S2D1 alone. The negative (placebo) control was Al(OH).sub.3-adjuvant alone. All antigens were formulated in PBS with aluminium hydroxide (Al(OH).sub.3) at a final concentration of 0.15%.

[0429] *(Chimeric OspA ST1/ST2 (SEQ ID NO: 212) is an OspA chimera consisting of the first 10 amino acids of the N-terminal portion of OspB (strain B31), amino acids 11-200 of serotype 1 OspA, fused with the last 201-255 amino acids from the C-terminal portion of serotype 2 OspA and wherein the hLFA-1-like sequence of the serotype 1 OspA (146-170) is replaced with the homologous sequence from a serotype 2 OspA The serotype 2 OspA sequence is followed by two amino acids which are added because of the cloning site (XhoI) ahead of the stop codon in the vector.)

[0430] One week after the third immunization, blood was collected from the facial vein and hyper-immune sera were prepared. All animal experiments were conducted in accordance with Austrian law (BGB1 Nr. 501/1989) and approved by Magistratsabteilung 58.

[0431] Needle Challenge of Immunized Mice with In Vitro Grown Borrelia

[0432] Two weeks after the last immunization, the mice were challenged s.c. with Borrelia spirochetes diluted in 100 L growth medium (BSKII). The challenge doses were strain-dependent, the virulence of the individual strains were assessed by challenge experiments for determination of ID.sub.50. Doses employed for needle challenge experiments ranged from 20 to 50 times the ID.sub.50. Four weeks after needle challenge, mice were sacrificed and blood and tissues were collected for readout methods to determine the infection status.

[0433] Tick Challenge of Immunized Mice and Collection of Sera and Tissues (Herein Referred to Also as Tick Challenge Method)

[0434] In order to challenge the immunized mice with B. afzelii, the hair of the back of each mouse was removed with Veet Cream (Reckitt Benckiser, United Kingdom) and a small ventilated container was glued to the skin with super glue (Pattex, Germany). Thereafter, one or two I. ricinus nymphs infected with B. afzelii, strain IS1, were applied per mouse, allowed to attach and feed until they are fully engorged and drop off. The feeding status was monitored for each individual tick and only mice where at least one fully-fed tick was collected were included in the final readout.

[0435] Results

[0436] Lipidated mutant OspA fragment heterodimers that were not His-tagged were combined at a 1:1:1 ratio and tested for protective capacity against Borrelia challenge Immunized mice were challenged with B. afzelii (ST2, strain IS1, tick challenge) or with B. burgdorferi s.s. (ST1, strain ZS7, needle challenge) in two experiments each. Other OspA-based antigens included Lip-S1D1-S2D2 in all four experiments and Lip-OspA1-His and lipidated chimeric OspA ST1/ST2 in Experiments 15 and 16. A group of mice immunized with Al(OH).sub.3 adjuvant alone served as a negative control group in each experiment. For challenge with ticks, 1-2 ticks were applied per mouse and only mice from which at least one tick fed until fully engorged were included in the final readout. However, no distinction was made between mice from which one or two fully fed ticks were collected. The protection data from the four experiments are summarized in Table 5.

[0437] The combination vaccine containing three lipidated mutant OspA fragment heterodimers at a 1:1:1 ratio conferred statistically-significant protection (Fisher's exact test, two-tailed) in all four challenge experiments as compared to the negative control group. The infectious status of each mouse was determined using VlsE ELISA in combination with recA qPCR. A mouse was regarded as infected when at least one method gave a positive result.

Example 6 Binding of Antibodies from the Sera of Mice Immunized with Mutant OspA Fragment Heterodimers to the Cell Surface of Borrelia

[0438] Experimental Procedures

[0439] Immunization of Mice

[0440] Female C3H/HeN mice were used for all studies. Prior to each challenge, groups of twenty 8-week-old mice were bled via the facial vein and pre-immune sera were prepared and pooled. Three s.c. immunizations of 100 L each were administered at two week intervals. Each dose contained 1 g of each of the respective proteins: Lip-S1D1-S2D1, Lip-S4D1-S3D1 and Lip-S5D1-S6D1 (combination vaccine), or 1 g lipidated full-length OspA protein (ST1-ST6 as indicated) or 1 g OspA heterodimer alone (Lip-SID1-S2D1, Lip-S4D1-S3D1 or Lip-S5D1-S6D1, as indicated) adjuvanted with aluminium hydroxide at a final concentration of 0.15%. The negative (placebo) control was Al(OH).sub.3 adjuvant alone. One week after the third immunization, blood was collected from the facial vein and hyper-immune sera were prepared. All animal experiments were conducted in accordance with Austrian law (BGB1 Nr. 501/1989) and approved by Magistratsabteilung 58.

[0441] Flow Cytometry to Assess Binding to Borrelia

[0442] Spirochetes (110.sup.6) were mixed with an equal volume of 4% paraformaldehyde and incubated for 2 hours at room temperature in a 96-well plate (Nunclon 96U, Nunc). The plate was centrifuged for 5 minutes at 2,000 g and the supernatant was discarded. Cells were washed with 150 L HBSS with 2% BSA (HBSS-B), centrifuged as above and the supernatant was discarded. Mouse sera were heat inactivated by incubating them at 56 C. for 35 minutes. Heat-inactivated sera were diluted in HBSS-B and sterile filtered by centrifuging 4,000 g for 3 minutes using Costar spin-X centrifuge tube filters (0.22 m, Corning, USA). Spirochetes were dissolved in 100 I., serum and incubated for 45 minutes at room temperature. The plate was centrifuged for 15 minutes at 2,000 g and the supernatant was discarded. The cells were washed once with 150 L HBSS-B and then dissolved in 100 L HBSS-B.

[0443] One microliter secondary antibody (PE conjugated goat anti-mouse IgG, Beckman Coulter, USA) was added to the cells and incubated at room temperature for 45 minutes in the dark. Spirochetes were washed once with 150 L HBSS-B and then dissolved in 200 L HBSS containing 2.5 M SYTO-17 DNA dye and incubated for 10 minutes at room temperature in the dark. The stained spirochetes were pelleted by centrifuging for 5 minutes at 2000 g and subsequently dissolved in 200 L HBSS. Labelled spirochetes were measured with a FC500 (Beckman Coulter) flow cytometer, gated for SYTO-17 positive events. Values obtained with sera from the placebo-immunized group were subtracted from the values observed with sera from the heterodimer-immunized groups to control for non-specific binding.

[0444] Results

[0445] Binding of antibodies from hyperimmune mouse sera was observed in the case of different Borreliae expressing all six OspA serotypes, indicating that the antibodies generated in response to all of the antigens are functionally active and can bind native OspA in situ. The fluorescence intensity was linear over a large range of serum dilutions. For most OspA serotypes, the observed fluorescence intensity with heterodimer-generated sera was comparable to the fluorescence intensity seen with sera generated with lipidated full-length OspA.

Example 7 Formulation Studies

[0446] Studies regarding the formulation of the combination vaccine of the invention were carried out in order to optimize stability. Different types of buffers and stabilizers were tested at various concentrations in combination with aluminum hydroxide and antigen. An optimal formulation of 40 g/mL each of three heterodimers (120 g total protein), 10 mM sodium phosphate, 150 mM sodium chloride, 10 mM L-Methionine, 5% Sucrose, 0.05% Tween 20 (polysorbate 20) and 0.15% (w/v) aluminium hydroxide at pH 6.70.2 was determined.#

TABLE-US-00012 SEQUENCES SEQIDNO:1 S2D0-His:aminoacidsofpositions131-273ofBorreliaafzeliistrainK78,OspAserotype2,wild-type sequence,C-terminalHistag(GLEHHHHHH) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LKNALKGLEHHHHHH SEQIDNO:2 S2D1-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype1(aa 182and269),C-terminalHistag(GLEHHHHHH) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKCGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LCNALKGLEHHHHHH SEQIDNO:3 S2D2-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype2(aa 182and272),C-terminalHistag(GLEHHHHHH) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKCGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LKNACKGLEHHHHHH SEQIDNO:4 S2D3-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype3(aa 244and259),C-terminalHistag(GLEHHHHHH) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTICVQKYDSAGTNLEGTCVEIKTLDE LKNALKGLEHHHHHH SEQIDNO:5 S2D4-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype4(aa 141and241),C-terminalHistag(GLEHHHHHH) ELSAKTMTRECGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQCTITVQKYDSAGTNLEGTAVEIKTLDE LKNALKGLEHHHHHH SEQIDNO:6 S2D5-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype5(aa 165and265),C-terminalHistag(GLEHHHHHH) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNCTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTCDE LKNALKGLEHHHHHH SEQIDNO:7 S2D6-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype6(aa 185and272),C-terminalHistag(GLEHHHHHH) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTCTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LKNACKGLEHHHHHH SEQIDNO:8 S2D7-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype7(aa 199and223),C-terminalHistag(GLEHHHHHH) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT CALNDINTTQATKKTGAWDSKTSTCTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LKNALKGLEHHHHHH SEQIDNO:9 S2D8-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype8(aa 243and262),C-terminalHistag(GLEHHHHHH) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTCTVQKYDSAGTNLEGTAVECKTLD ELKNALKGLEHHHHHH SEQIDNO:10 S2D9-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype9(aa 184and204),C-terminalHistag(GLEHHHHHH) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGCVTLSKEIAKSGEVT VALNDCNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LKNALKGLEHHHHHH SEQIDNO:11 S2D10-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype10(aa 201and214),C-terminalHistag(GLEHHHHHH) ELSAKTMTRENGTKLEYTEMKSDGIGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VACNDTNTTQATKKTCAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LKNALKGLEHHHHHH SEQIDNO:12 S2D11-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype11(aa 246and259),C-terminalHistag(GLEHHHHHH) ELSAKTMTRENGTKLEYTEMKSDGIGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVCKYDSAGTNLEGTCVEIKTLDE LKNALKGLEHHHHHH SEQIDNO:13 S2D12-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype12(aa 167and178),C-terminalHistag(GLEHHHHHH) ELSAKTMTRENGTKLEYTEMKSDGIGKAKEVLKNFTCEGKVANDKVTCEVKEGTVTLSKEIAKSGEV TVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLD ELKNALKGLEHHHHHH SEQIDNO:14 BorreliaOspAlipidationsignal MKKYLLGIGLILALIA SEQIDNO:15 BorreliaOspBlipidationsignal MRLLIGFALALALIG SEQIDNO:16 E.coliIpplipidationsignal MKATKLVLGAVILGSTLLAG SEQIDNO:17 hLFA-1-likesequencefromB.burgdorferis.s.strainB31 GYVLEGTLTAE SEQIDNO:18 Non-hLFA-1-likesequencefromB.afzeliistrainK78 NFTLEGKVAND SEQIDNO:19 B.afzelii(strainK78;OspAserotype2) MKKYLLGIGLILALIACKQNVSSLDEKNSASVDLPGEMKVLVSKEKDKDGKYSLKATVDKIELKGTSDK DNGSGVLEGTKDDKSKAKLTIADDLSKTTFELFKEDGKTLVSRKVSSKDKTSTDEMFNEKGELSAKT MTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVTVALNDT NTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDELKNALK SEQIDNO:20 B.burgdorferis.s.(strainB31,OspAserotype1) MKKYLLGIGLILALIACKQNVSSLDEKNSVSVDLPGEMKVLVSKEKNKDGKYDLIATVDKLELKGTSDK NNGSGVLEGVKADKSKVKLTISDDLGQTTLEVFKEDGKTLVSKKVTSKDKSSTEEKFNEKGEVSEKIIT RADGTRLEYTGIKSDGSGKAKEVLKGYVLEGTLTAEKTTLVVKEGTVTLSKNISKSGEVSVELNDTDS SAATKKTAAWNSGTSTLTITVNSKKTKDLVFTKENTITVQQYDSNGTKLEGSAVEITKLDEIKNALK SEQIDNO:21 B.garinii(strainPBr,OspAserotype3) MKKYLLGIGLILALIACKQNVSSLDEKNSVSVDLPGGMKVLVSKEKDKDGKYSLMATVEKLELKGTSD KSNGSGVLEGEKADKSKAKLTISQDLNQTTFEIFKEDGKTLVSRKVNSKDKSSTEEKFNDKGKLSEKV VTRANGTRLEYTEIKNDGSGKAKEVLKGFALEGTLTDGGETKLTVTEGTVTLSKNISKSGEITVALNDT ETTPADKKTGEWKSDTSTLTISKNSQKPKQLVFTKENTITVQNYNRAGNALEGSPAEIKDLAELKAALK SEQIDNO:22 B.bavariensis(strainPBi,OspAserotype4) MKKYLLGIGLILALIACKQNVSSLDEKNSVSVDLPGEMKVLVSKEKDKDGKYSLMATVDKLELKGTSD KSNGSGTLEGEKSDKSKAKLTISEDLSKTTFEIFKEDGKTLVSKKVNSKDKSSIEEKFNAKGELSEKTIL RANGTRLEYTEIKSDGTGKAKEVLKDFALEGTLAADKTTLKVTEGTVVLSKHIPNSGEITVELNDSNST QATKKTGKWDSNTSTLTISVNSKKTKNIVFTKEDTITVOKYDSAGTNLEGNAVEIKTLDELKNALK SEQIDNO:23 B.garinii(strainPHei,OspAserotype5) MKKYLLGIGLILALIACKQNVSSLDEKNSVSVDLPGGMKVLVSKEKDKDGKYSLMATVEKLELKGTSD KNNGSGTLEGEKTDKSKVKLTIAEDLSKTTFEIFKEDGKTLVSKKVTLKDKSSTEEKFNEKGEISEKTIV RANGTRLEYTDIKSDKTGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVTLSKNISKSGEITVALDDTDS SGNKKSGTWDSGTSTLTISKNRTKTKQLVFTKEDTITVQNYDSAGTNLEGKAVEITTLKELKNALK SEQIDNO:24 B.garinii(strainDK29,OspAserotype6) MKKYLLGIGLILALIACKQNVSSLDEKNSVSVDLPGGMTVLVSKEKDKDGKYSLEATVDKLELKGTSDK NNGSGTLEGEKTDKSKVKSTIADDLSQTKFEIFKEDGKTLVSKKVTLKDKSSTEEKFNGKGETSEKTIV RANGTRLEYTDIKSDGSGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVVLSKNILKSGEITAALDDSDT TRATKKTGKWDSKTSTLTISVNSQKTKNLVFTKEDTITVQRYDSAGTNLEGKAVEITTLKELKNALK SEQIDNO:25 B.garinii(strainT25,OspAserotype7) MKKYLLGIGLILALIACKQNVSSLDEKNSVSVDLPGEMKVLVSKEKDKDGKYSLEATVDKLELKGTSDK NNGSGVLEGVKAAKSKAKLTIADDLSQTKFEIFKEDGKTLVSKKVTLKDKSSTEEKFNDKGKLSEKVV TRANGTRLEYTEIQNDGSGKAKEVLKSLTLEGTLTADGETKLTVEAGTVTLSKNISESGEITVELKDTE TTPADKKSGTWDSKTSTLTISKNSQKTKQLVFTKENTITVQKYNTAGTKLEGSPAEIKDLEALKAALK SEQIDNO:26 ForwardPrimer GTATGTTTAGTGAGGGGGGTG SEQIDNO:27 ReversePrimer GGATCATAGCTCAGGTGGTTAG SEQIDNO:28 ForwardNestedPrimer AGGGGGGTGAAGTCGTAACAAG SEQIDNO:29 ReversedNestedPrimer GTCTGATAAACCTGAGGTCGGA SEQIDNO:30 25-merpeptide MKKDDQIAAAMVLRGMAKDGQFALK SEQIDNO:31 Mousecathelin RLAGLLRKGGEKIGEKLKKIGQKIKNFFQKLVPQPE SEQIDNO:32 5-(dIdC).sub.13-3 dIdCdIdCdIdCdIdCdIdCdIdCdIdCdIdCdIdCdIdCdIdCdIdCdIdC SEQIDNO:33 KLKpeptide KLKLLLLLKLK SEQIDNO:34 B.afzelii(strainK78,serotype2),OspAaa126-273 FNEKGELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAK SGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEI KTLDELKNALK SEQIDNO:35 B.afzelii(strainK78,serotype2),OspAaa131-273 ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LKNALK SEQIDNO:36 peptidelinker GGGGGGGG SEQIDNO:37 peptidelinker GGGGGGGGGGGG SEQIDNO:38 peptidelinker GAGA SEQIDNO:39 peptidelinker GAGAGAGA SEQIDNO:40 peptidelinker GAGAGAGAGAGA SEQIDNO:41 peptidelinker GGGSGGGS SEQIDNO:42 peptidelinker GGGSGGGSGGGS SEQIDNO:43 S1D4-S2D4_aa:HeterodimerfusionproteinofOspAserotypes1and2bothwithdisulfidebondtype 4,LN1linkersequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequence NFTLEGKVAND FNEKGEVSEKIITRACGTRLEYTGIKSDGSGKAKEVLKNFTLEGKVANDKTTLVVKEGTVTLSKNISKS GEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSKKTKDLVFTKECTITVQQYDSNGTKLEGSAVEIT KLDEIKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGELSAKTMTRECGTKLEYTEMKSDGTGKAKE VLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNS KKTTQLVFTKQCTITVQKYDSAGTNLEGTAVEIKTLDELKNALK SEQIDNO:44 Lip-S1D4-52D4_nt:CodingsequenceforfusionproteinsofOspAserotypes1and2bothwith disulfidebondtype4,E.coliIpplipidationsignal,LN1linkersequence,aa164-174ofOspAserotype 1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAGTCTCGGAAAAAATCATTACCCGTGCTTGCGGCACCCGTCT GGAATACACCGGCATTAAATCGGATGGCAGCGGCAAAGCGAAGGAAGTTCTGAAAAACTTTACC CTGGAAGGCAAAGTCGCAAATGATAAGACCACCCTGGTGGTGAAAGAAGGCACCGTTACGCTGA GCAAAAACATTAGTAAGTCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGC CACCAAAAAGACGGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATTCCAAA AAGACCAAAGATCTGGTCTTCACGAAAGAATGCACCATCACGGTGCAGCAATATGACAGCAACG GTACCAAACTGGAAGGCTCTGCGGTGGAAATCACGAAACTGGATGAAATCAAAAATGCTCTGAAA GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAATGCGGCACCAAACTGGAATAT ACGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCCTGGAAG GCAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAAGAAGGCACCGTTACGCTGTCAAAAGA AATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCGACCAAG AAAACCGGCGCCIGGGACAGCAAAACCTCTACGCTGACCATTAGIGTTAACAGCAAGAAAACCA CGCAGCTGGTCTTCACCAAACAATGTACGATCACCGTGCAGAAATACGATAGTGCGGGTACCAA CCTGGAAGGCACCGCTGTTGAAATCAAAACCCTGGACGAACTGAAAAACGCCCTGAAA SEQIDNO:45 Lip-S1D4-S2D4_His_aa:HeterodimerfusionproteinofOspAserotypes1and2bothwithdisulfide bondtype4,N-terminalCSSforadditionoflipids,N-terminallipidation,LN1linkersequence,aa164- 174ofOspAserotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND,C-terminalHistag (GLEHHHHHH) LipCSSFNEKGEVSEKIITRACGTRLEYTGIKSDGSGKAKEVLKNFTLEGKVANDKTTLVVKEGTVTLSK NISKSGEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSKKTKDLVFTKECTITVQQYDSNGTKLEG SAVEITKLDEIKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGELSAKTMTRECGTKLEYTEMKSDGT GKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTL TISVNSKKTTQLVFTKQCTITVQKYDSAGTNLEGTAVEIKTLDELKNALKGLEHHHHHH SEQIDNO:46 Lip-S1D4-S2D4_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotypes1and2 bothwithdisulfidebondtype4,E.coliIpplipidationsignal,LN1linkersequence,aa164-174ofOspA serotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND,C-terminalHistag (GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAGTCTCGGAAAAAATCATTACCCGTGCTTGCGGCACCCGTCT GGAATACACCGGCATTAAATCGGATGGCAGCGGCAAAGCGAAGGAAGTTCTGAAAAACTTTACC CTGGAAGGCAAAGTCGCAAATGATAAGACCACCCTGGTGGTGAAAGAAGGCACCGTTACGCTGA GCAAAAACATTAGTAAGTCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGC CACCAAAAAGACGGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATTCCAAA AAGACCAAAGATCTGGTCTTCACGAAAGAATGCACCATCACGGTGCAGCAATATGACAGCAACG GTACCAAACTGGAAGGCTCTGCGGTGGAAATCACGAAACTGGATGAAATCAAAAATGCTCTGAAA GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAATGCGGCACCAAACTGGAATAT ACGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCCTGGAAG GCAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAAGAAGGCACCGTTACGCTGTCAAAAGA AATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCGACCAAG AAAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAACAGCAAGAAAACCA CGCAGCTGGTCTTCACCAAACAATGTACGATCACCGTGCAGAAATACGATAGTGCGGGTACCAA CCTGGAAGGCACCGCTGTTGAAATCAAAACCCTGGACGAACTGAAAAACGCCCTGAAAGGCCTC GAGCACCACCACCACCACCAC SEQIDNO:47 S1D1-S2D1_aa:HeterodimerfusionproteinofOspAserotype1andOspAserotype2withdisulfide bondtype1,LN1linkersequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-like sequenceNFTLEGKVAND FNEKGEVSEKIITRADGTRLEYTGIKSDGSGKAKEVLKNFTLEGKVANDKTTLVVKCGTVTLSKNISKS GEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSKKTKDLVFTKENTITVQQYDSNGTKLEGSAVEIT KLDEICNALKGTSDKNNGSGSKEKNKDGKYSFNEKGELSAKTMTRENGTKLEYTEMKSDGTGKAKE VLKNFTLEGKVANDKVTLEVKCGTVTLSKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNS KKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDELCNALK SEQIDNO:48 Lip-S1D1-S2D1_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotype1andOspAserotype2withdisulfidebondtype1,E.coliIpplipidationsignal,LN1linker sequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAGTCAGCGAAAAAATCATTACCCGCGCAGACGGCACCCGCCT GGAATACACCGGCATCAAATCGGACGGCAGCGGCAAAGCGAAAGAAGTTCTGAAAAACTTTACC CTGGAAGGCAAAGTCGCAAATGATAAAACCACCCTGGTGGTGAAATGCGGCACCGTTACGCTGA GCAAAAACATTAGTAAATCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGC CACCAAGAAAACCGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATAGCAAG AAAACCAAAGATCTGGTCTTCACGAAAGAAAACACCATCACGGTGCAGCAATATGACAGCAATGG TACCAAACTGGAAGGCTCCGCTGTGGAAATCACGAAACTGGATGAAATCTGTAATGCTCTGAAAG GTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATTC AACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAAAACGGCACCAAACTGGAATATA CGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCCTGGAAGG CAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAATGCGGCACCGTTACGCTGTCAAAAGAA ATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCGACCAAGA AAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAATAGCAAGAAAACCAC GCAGCTGGTCTTCACCAAACAAGATACGATCACCGTGCAGAAATACGACAGTGCGGGTACCAAC CTGGAAGGCACGGCTGTTGAAATCAAAACCCTGGACGAACTGTGTAACGCCCTGAAA SEQIDNO:49 Lip-S1D1-S2D1_His_aa:HeterodimerfusionproteinofOspAserotype1andOspAserotype2with disulfidebondtype1,N-terminalCSSforadditionoflipids,N-terminallipidation,LN1linker sequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND,C-terminal Histag(GLEHHHHHH) LipCSSFNEKGEVSEKIITRADGTRLEYTGIKSDGSGKAKEVLKNFTLEGKVANDKTTLVVKCGTVTLSK NISKSGEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSKKTKDLVFTKENTITVQQYDSNGTKLEG SAVEITKLDEICNALKGTSDKNNGSGSKEKNKDGKYSFNEKGELSAKTMTRENGTKLEYTEMKSDGT GKAKEVLKNFTLEGKVANDKVTLEVKCGTVTLSKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTL TISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDELCNALKGLEHHHHHH SEQIDNO:50 Lip-S1D1-S2D1_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotype1and OspAserotype2withdisulfidebondtype1,E.coliIpplipidationsignal,LN1linkersequence,aa164- 174ofOspAserotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND,C-terminalHistag (GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAGTCAGCGAAAAAATCATTACCCGCGCAGACGGCACCCGCCT GGAATACACCGGCATCAAATCGGACGGCAGCGGCAAAGCGAAAGAAGTTCTGAAAAACTTTACC CTGGAAGGCAAAGTCGCAAATGATAAAACCACCCTGGTGGTGAAATGCGGCACCGTTACGCTGA GCAAAAACATTAGTAAATCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGC CACCAAGAAAACCGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATAGCAAG AAAACCAAAGATCTGGTCTTCACGAAAGAAAACACCATCACGGTGCAGCAATATGACAGCAATGG TACCAAACTGGAAGGCTCCGCTGTGGAAATCACGAAACTGGATGAAATCTGTAATGCTCTGAAAG GTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATTC AACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAAAACGGCACCAAACTGGAATATA CGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCCTGGAAGG CAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAATGCGGCACCGTTACGCTGTCAAAAGAA ATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCGACCAAGA AAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAATAGCAAGAAAACCAC GCAGCTGGTCTTCACCAAACAAGATACGATCACCGTGCAGAAATACGACAGTGCGGGTACCAAC CTGGAAGGCACGGCTGTTGAAATCAAAACCCTGGACGAACTGTGTAACGCCCTGAAAGGCCTCG AGCACCACCACCACCACCAC SEQIDNO:51 S3D4-S4D4_aa:HeterodimerfusionproteinofOspAserotype3andOspAserotype4withdisulfide bondtype4,LN1linkersequence FNEKGKLSEKVVTRACGTRLEYTEIKNDGSGKAKEVLKGFALEGTLTDGGETKLTVTEGTVTLSKNIS KSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNSQKPKQLVFTKECTITVQNYNRAGNALEGSPA EIKDLAELKAALKGTSDKNNGSGSKEKNKDGKYSFNAKGELSEKTILRACGTRLEYTEIKSDGTGKAK EVLKDFALEGTLAADKTTLKVTEGTVVLSKHIPNSGEITVELNDSNSTQATKKTGKWDSNTSTLTISVN SKKTKNIVFTKECTITVQKYDSAGTNLEGNAVEIKTLDELKNALK SEQIDNO:52 Lip-S3D4-S4D4_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotype3andOspAserotype4withdisulfidebondtype4,E.coliIpplipidationsignal,LN1linker sequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCAAACTGTCAGAAAAAGTGGTCACCCGCGCTTGTGGCACCCGCCT GGAATACACCGAAATCAAAAACGACGGCTCGGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCC CTGGAAGGTACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGGAAGGCACCGTTACG CTGTCTAAAAACATTAGCAAGTCTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCC GGCTGACAAAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCG CAGAAACCGAAGCAACTGGTCTTCACCAAAGAATGCACGATCACCGTGCAGAACTATAATCGTG CCGGTAATGCTCTGGAAGGCTCCCCGGCTGAAATCAAGGACCTGGCGGAACTGAAGGCGGCAC TGAAAGGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTA CTCATTCAACGCTAAAGGTGAACTGTCGGAAAAAACCATCCTGCGCGCCTGTGGCACCCGCCTG GAATACACGGAAATCAAGTCGGACGGCACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCTC TGGAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTTCTGA GCAAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGCG ACCAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAAAA GACCAAAAATATTGTGTTCACGAAGGAATGCACGATCACCGTTCAAAAATATGATTCCGCAGGTA CCAACCTGGAAGGCAACGCTGTGGAAATCAAAACCCTGGACGAACTGAAAAATGCTCTGAAG SEQIDNO:53 Lip-S3D4-S4D4_His_aa:HeterodimerfusionproteinofOspAserotype3andOspAserotype4with disulfidebondtype4,N-terminalCSSforadditionoflipids,N-terminallipidation,LN1linker sequence,C-terminalHistag(GLEHHHHHH) LipCSSFNEKGKLSEKVVTRACGTRLEYTEIKNDGSGKAKEVLKGFALEGTLTDGGETKLTVTEGTVTL SKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNSQKPKQLVFTKECTITVQNYNRAGNALE GSPAEIKDLAELKAALKGTSDKNNGSGSKEKNKDGKYSFNAKGELSEKTILRACGTRLEYTEIKSDGT GKAKEVLKDFALEGTLAADKTTLKVTEGTVVLSKHIPNSGEITVELNDSNSTQATKKTGKWDSNTSTL TISVNSKKTKNIVFTKECTITVQKYDSAGTNLEGNAVEIKTLDELKNALKGLEHHHHHH SEQIDNO:54 Lip-S3D4-S4D4_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotype3and OspAserotype4withdisulfidebondtype4,E.coliIpplipidationsignal,N-terminalCSSfor additionoflipids,LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCAAACTGTCAGAAAAAGTGGTCACCCGCGCTTGTGGCACCCGCCT GGAATACACCGAAATCAAAAACGACGGCTCGGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCC CTGGAAGGTACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGGAAGGCACCGTTACG CTGTCTAAAAACATTAGCAAGTCTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCC GGCTGACAAAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCG CAGAAACCGAAGCAACTGGTCTTCACCAAAGAATGCACGATCACCGTGCAGAACTATAATCGTG CCGGTAATGCTCTGGAAGGCTCCCCGGCTGAAATCAAGGACCTGGCGGAACTGAAGGCGGCAC TGAAAGGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTA CTCATTCAACGCTAAAGGTGAACTGTCGGAAAAAACCATCCTGCGCGCCTGTGGCACCCGCCTG GAATACACGGAAATCAAGTCGGACGGCACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCTC TGGAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTTCTGA GCAAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGCG ACCAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAAAA GACCAAAAATATTGTGTTCACGAAGGAATGCACGATCACCGTTCAAAAATATGATTCCGCAGGTA CCAACCTGGAAGGCAACGCTGTGGAAATCAAAACCCTGGACGAACTGAAAAATGCTCTGAAGGG TCTCGAGCACCACCACCACCACCAC SEQIDNO:55 S3D1-S4D1_aa:HeterodimerfusionproteinofOspAserotypes3and4bothwithdisulfidebondtype 1,LN1linkersequence FNEKGKLSEKVVTRANGTRLEYTEIKNDGSGKAKEVLKGFALEGTLTDGGETKLTVTCGTVTLSKNIS KSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNSQKPKOLVFTKENTITVONYNRAGNALEGSPA EIKDLAELCAALKGTSDKNNGSGSKEKNKDGKYSFNAKGELSEKTILRANGTRLEYTEIKSDGTGKAK EVLKDFALEGTLAADKTTLKVTCGTVVLSKHIPNSGEITVELNDSNSTQATKKTGKWDSNTSTLTISVN SKKTKNIVFTKEDTITVQKYDSAGTNLEGNAVEIKTLDELCNALK SEQIDNO:56 Lip-S3D1-S4D1_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotypes3and4bothwithdisulfidebondtype1,E.coliIpplipidationsignal,N-terminalCSSfor additionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCAAACTGTCGGAAAAAGTGGTCACCCGCGCAAATGGCACCCGCCT GGAATACACGGAAATCAAAAACGATGGTAGCGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCC CTGGAAGGTACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGTGCGGCACCGTTACG CTGTCTAAAAACATTAGCAAGICTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCC GGCTGACAAAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCG CAGAAACCGAAGCAACTGGTCTTCACCAAAGAAAACACGATCACCGTGCAGAACTATAATCGTGC CGGTAATGCTCTGGAAGGCTCACCGGCTGAAATCAAGGACCTGGCTGAACTGTGTGCGGCACT GAAAGGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTAC TCATTCAACGCTAAAGGTGAACTGAGCGAAAAAACGATCCTGCGTGCGAATGGCACCCGTCTGG AATACACCGAAATCAAATCCGATGGTACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCTCTG GAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGTTCTGAGC AAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGCAAC CAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAAAAGA CCAAAAATATTGTGTTCACGAAGGAAGATACGATCACCGTTCAAAAATATGACTCCGCGGGCACC AACCTGGAAGGCAATGCCGTCGAAATCAAAACCCTGGATGAACTGTGTAATGCTCTGAAG SEQIDNO:57 Lip-S3D1-S4D1_His_aa:HeterodimerfusionproteinofOspAserotypes3and4bothwithdisulfide bondtype1,E.coliIpplipidationsignal,N-terminalCSSforadditionoflipids,N-terminal lipidation,LN1linkersequence,C-terminalHistag(GLEHHHHHH) LipCSSFNEKGKLSEKVVTRANGTRLEYTEIKNDGSGKAKEVLKGFALEGTLTDGGETKLTVTCGTVTL SKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNSQKPKQLVFTKENTITVQNYNRAGNALE GSPAEIKDLAELCAALKGTSDKNNGSGSKEKNKDGKYSFNAKGELSEKTILRANGTRLEYTEIKSDGT GKAKEVLKDFALEGTLAADKTTLKVTCGTVVLSKHIPNSGEITVELNDSNSTQATKKTGKWDSNTSTL TISVNSKKTKNIVFTKEDTITVQKYDSAGTNLEGNAVEIKTLDELCNALKGLEHHHHHH SEQIDNO:58 Lip-S3D1-S4D1_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotypes3and4 bothwithdisulfidebondtype1,E.coliIpplipidationsignal,N-terminalCSSforadditionof lipids,LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCAAACTGTCGGAAAAAGTGGTCACCCGCGCAAATGGCACCCGCCT GGAATACACGGAAATCAAAAACGATGGTAGCGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCC CTGGAAGGTACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGTGCGGCACCGTTACG CTGTCTAAAAACATTAGCAAGTCTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCC GGCTGACAAAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCG CAGAAACCGAAGCAACTGGTCTTCACCAAAGAAAACACGATCACCGTGCAGAACTATAATCGTGC CGGTAATGCTCTGGAAGGCTCACCGGCTGAAATCAAGGACCTGGCTGAACTGTGTGCGGCACT GAAAGGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTAC TCATTCAACGCTAAAGGTGAACTGAGCGAAAAAACGATCCTGCGTGCGAATGGCACCCGTCTGG AATACACCGAAATCAAATCCGATGGTACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCTCTG GAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGTTCTGAGC AAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGCAAC CAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAAAAGA CCAAAAATATTGTGTTCACGAAGGAAGATACGATCACCGTTCAAAAATATGACTCCGCGGGCACC AACCTGGAAGGCAATGCCGTCGAAATCAAAACCCTGGATGAACTGTGTAATGCTCTGAAGGGTC TCGAGCACCACCACCACCACCAC SEQIDNO:59 S5D4-S6D4_aa:HeterodimerfusionproteinOspAserotypes5and6bothwithdisulfidebondtype4, LN1linkersequence FNEKGEISEKTIVRACGTRLEYTDIKSDKTGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVTLSKNISKS GEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRTKTKOLVFTKECTITVQNYDSAGTNLEGKAVEITT LKELKNALKGTSDKNNGSGSKEKNKDGKYSFNGKGETSEKTIVRACGTRLEYTDIKSDGSGKAKEVL KDFTLEGTLAADGKTTLKVTEGTVVLSKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQ KTKNLVFTKECTITVQRYDSAGTNLEGKAVEITTLKELKNALK SEQIDNO:60 Lip-S5D4-S6D4_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsOspA serotypes5and6bothwithdisulfidebondtype4,E.coliIpplipidationsignal,N-terminalCSSfor additionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAATCAGTGAAAAAACCATTGTGCGTGCGTGTGGCACCCGTCT GGAATATACCGACATCAAGAGCGATAAAACGGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGC TGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCGAAGGTACCGTTACGC TGTCCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCTGGATGACACCGATAGCTCTGG CAACAAAAAGAGCGGTACCTGGGACTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACG AAAACCAAGCAGCTGGTCTTCACGAAAGAATGCACGATCACCGTGCAAAACTATGATAGCGCAG GTACCAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGAAGAATGCTCTGAAA GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGGCAAAGGTGAAACGAGTGAAAAAACGATTGTTCGCGCCTGTGGCACCCGCCTGGAATAC ACGGATATCAAGTCGGATGGTTCGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTACGCTGGAAG GTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTTCTGTCAA AAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACGCGTGCTAC GAAAAAGACCGGTAAATGGGACAGCAAGACCTCTACGCTGACCATTAGTGTCAACTCCCAGAAA ACGAAGAATCTGGTGTTCACCAAAGAATGCACGATCACCGTTCAACGCTATGATAGTGCGGGCA CCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGAAGAATGCTCTGAAA SEQIDNO:61 Lip-S5D4-S6D4_His_aa:HeterodimerfusionproteinOspAserotypesSand6bothwithdisulfidebond type4,N-terminalCSSforadditionoflipids,N-terminallipidation,LN1linkersequence,C-terminal Histag(GLEHHHHHH) LipCSSFNEKGEISEKTIVRACGTRLEYTDIKSDKTGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVTLS KNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRTKTKOLVFTKECTITVQNYDSAGTNLEGK AVEITTLKELKNALKGTSDKNNGSGSKEKNKDGKYSFNGKGETSEKTIVRACGTRLEYTDIKSDGSGK AKEVLKDFTLEGTLAADGKTTLKVTEGTVVLSKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTIS VNSQKTKNLVFTKECTITVORYDSAGTNLEGKAVEITTLKELKNALKGLEHHHHHH SEQIDNO:62 Lip-S5D4-S6D4_His_nt:CodingsequenceforheterodimerfusionproteinOspAserotypes5and6 bothwithdisulfidebondtype4,E.coliIpplipidationsignal,N-terminalCSSforadditionoflipids, LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAATCAGTGAAAAAACCATTGTGCGTGCGTGTGGCACCCGTCT GGAATATACCGACATCAAGAGCGATAAAACGGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGC TGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCGAAGGTACCGTTACGC TGTCCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCTGGATGACACCGATAGCTCTGG CAACAAAAAGAGCGGTACCTGGGACTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACG AAAACCAAGCAGCTGGTCTTCACGAAAGAATGCACGATCACCGTGCAAAACTATGATAGCGCAG GTACCAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGAAGAATGCTCTGAAA GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGGCAAAGGTGAAACGAGTGAAAAAACGATTGTTCGCGCCTGTGGCACCCGCCTGGAATAC ACGGATATCAAGTCGGATGGTTCGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTACGCTGGAAG GTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTTCTGTCAA AAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACGCGTGCTAC GAAAAAGACCGGTAAATGGGACAGCAAGACCTCTACGCTGACCATTAGTGTCAACTCCCAGAAA ACGAAGAATCTGGTGTTCACCAAAGAATGCACGATCACCGTTCAACGCTATGATAGTGCGGGCA CCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGAAGAATGCTCTGAAAGG TCTCGAGCACCACCACCACCACCAC SEQIDNO:63 S5D1-S6D1_aa:HeterodimerfusionproteinofOspAserotypes6bothwithdisulfidebondtype1,LN1 linkersequence FNEKGEISEKTIVRANGTRLEYTDIKSDKTGKAKEVLKDFTLEGTLAADGKTTLKVTCGTVTLSKNISKS GEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRTKTKQLVFTKEDTITVQNYDSAGTNLEGKAVEITT LKELCNALKGTSDKNNGSGSKEKNKDGKYSFNGKGETSEKTIVRANGTRLEYTDIKSDGSGKAKEVL KDFTLEGTLAADGKTTLKVTCGTVVLSKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQ KTKNLVFTKEDTITVQRYDSAGTNLEGKAVEITTLKELCNALK SEQIDNO:64 Lip-S5D1-S6D1_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotypes6bothwithdisulfidebondtype1,E.coliIpplipidationsignal,N-terminalCSSfor additionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAATCTCAGAAAAAACCATCGTCCGCGCTAACGGCACCCGCCT GGAATACACCGACATCAAATCAGACAAGACCGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGC TGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCTGCGGTACCGTTACGC TGTCCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCTGGATGACACCGATAGCTCTGG CAACAAAAAGAGCGGTACCTGGGATTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACG AAAACCAAGCAGCTGGTCTTCACGAAAGAAGATACGATCACCGTGCAAAACTATGACAGCGCAG GTACCAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGTGTAATGCTCTGAAA GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGGCAAAGGTGAAACGAGCGAAAAGACCATCGTGCGTGCGAACGGTACCCGCCTGGAATA TACGGACATTAAATCGGACGGCAGCGGCAAAGCAAAGGAAGTCCTGAAAGATTTTACGCTGGAA GGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGTTCTGTCA AAAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACGCGTGCTA CGAAAAAGACCGGTAAATGGGATAGCAAGACCTCTACGCTGACCATTAGTGTCAACTCCCAGAA AACGAAGAATCTGGTGTTCACCAAAGAAGATACGATCACCGTTCAACGCTATGACAGTGCGGGC ACCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGTGTAATGCTCTGAAA SEQIDNO:65 Lip-S5D1-S6D1_His_aa:HeterodimerfusionproteinofOspAserotypes6bothwithdisulfidebond type1,N-terminalCSSforadditionoflipids,N-terminallipidation,LN1linkersequence,C-terminal Histag(GLEHHHHHH) LipCSSFNEKGEISEKTIVRANGTRLEYTDIKSDKTGKAKEVLKDFTLEGTLAADGKTTLKVTCGTVTLS KNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRTKTKQLVFTKEDTITVQNYDSAGTNLEGK AVEITTLKELCNALKGTSDKNNGSGSKEKNKDGKYSFNGKGETSEKTIVRANGTRLEYTDIKSDGSGK AKEVLKDFTLEGTLAADGKTTLKVTCGTVVLSKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTIS VNSQKTKNLVFTKEDTITVQRYDSAGTNLEGKAVEITTLKELCNALKGLEHHHHHH SEQIDNO:66 Lip-S5D1-S6D1_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotypes6both withdisulfidebondtype1,E.coliIpplipidationsignal,N-terminalCSSforadditionoflipids, LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAATCTCAGAAAAAACCATCGTCCGCGCTAACGGCACCCGCCT GGAATACACCGACATCAAATCAGACAAGACCGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGC TGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCTGCGGTACCGTTACGC TGTCCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCTGGATGACACCGATAGCTCTGG CAACAAAAAGAGCGGTACCTGGGATTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACG AAAACCAAGCAGCTGGTCTTCACGAAAGAAGATACGATCACCGTGCAAAACTATGACAGCGCAG GTACCAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGTGTAATGCTCTGAAA GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGGCAAAGGTGAAACGAGCGAAAAGACCATCGTGCGTGCGAACGGTACCCGCCTGGAATA TACGGACATTAAATCGGACGGCAGCGGCAAAGCAAAGGAAGTCCTGAAAGATTTTACGCTGGAA GGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGTTCTGTCA AAAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACGCGTGCTA CGAAAAAGACCGGTAAATGGGATAGCAAGACCTCTACGCTGACCATTAGTGTCAACTCCCAGAA AACGAAGAATCTGGTGTTCACCAAAGAAGATACGATCACCGTTCAACGCTATGACAGTGCGGGC ACCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGTGTAATGCTCTGAAAG GTCTCGAGCACCACCACCACCACCAC SEQIDNO:67 S2D4-S1D4_aa:HeterodimerfusionproteinofOspAserotypes2and1bothwithdisulfidebondtype 4,LN1linkersequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequence NFTLEGKVAND FNEKGELSAKTMTRECGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAK SGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQCTITVQKYDSAGTNLEGTAVEI KTLDELKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEVSEKIITRACGTRLEYTGIKSDGSGKAKEV LKNFTLEGKVANDKTTLVVKEGTVTLSKNISKSGEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSK KTKDLVFTKECTITVQQYDSNGTKLEGSAVEITKLDEIKNALK SEQIDNO:68 Lip-S2D4-S1D4_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotypes2and1bothwithdisulfidebondtype4,E.coliIpplipidationsignal,N-terminalCSSfor additionoflipids,LN1linkersequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-like sequenceNFTLEGKVAND ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAATGCGGCACCAAACT GGAATATACGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCC TGGAAGGCAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAAGAAGGCACCGTTACGCTGTC AAAAGAAATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCG ACCAAGAAAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAACAGCAAGA AAACCACGCAGCTGGTCTTCACCAAACAATGTACGATCACCGTGCAGAAATACGATAGTGCGGG TACCAACCTGGAAGGCACCGCTGTTGAAATCAAAACCCTGGACGAACTGAAAAACGCCCTGAAA GGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGAAAAAGGCGAAGTCTCGGAAAAAATCATTACCCGTGCTTGCGGCACCCGTCTGGAATAC ACCGGCATTAAATCGGATGGCAGCGGCAAAGCGAAGGAAGTTCTGAAAAACTTTACCCTGGAAG GCAAAGTCGCAAATGATAAGACCACCCTGGTGGTGAAAGAAGGCACCGTTACGCTGAGCAAAAA CATTAGTAAGTCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGCCACCAAA AAGACGGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATTCCAAAAAGACCA AAGATCTGGTCTTCACGAAAGAATGCACCATCACGGTGCAGCAATATGACAGCAACGGTACCAA ACTGGAAGGCTCTGCGGTGGAAATCACGAAACTGGATGAAATCAAAAATGCACTGAAA SEQIDNO:69 Lip-S2D4-S1D4_His_aa:HeterodimerfusionproteinofOspAserotypes2and1bothwithdisulfide bondtype4,N-terminalCSSforadditionoflipids,N-terminallipidation,LN1linkersequence, aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND,C-terminalHistag (GLEHHHHHH) LipCSSFNEKGELSAKTMTRECGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTL SKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQCTITVQKYDSAGTNLE GTAVEIKTLDELKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEVSEKIITRACGTRLEYTGIKSDGS GKAKEVLKNFTLEGKVANDKTTLVVKEGTVTLSKNISKSGEVSVELNDTDSSAATKKTAAWNSGTSTL TITVNSKKTKDLVFTKECTITVQQYDSNGTKLEGSAVEITKLDEIKNALKGLEHHHHHH SEQIDNO:70 Lip-S2D4-S1D4_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotypes2and1 bothwithdisulfidebondtype4,E.coliIpplipidationsignal,N-terminalCSSforadditionof lipids,LN1linkersequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequence NFTLEGKVAND,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAATGCGGCACCAAACT GGAATATACGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCC TGGAAGGCAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAAGAAGGCACCGTTACGCTGTC AAAAGAAATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCG ACCAAGAAAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAACAGCAAGA AAACCACGCAGCTGGTCTTCACCAAACAATGTACGATCACCGTGCAGAAATACGATAGTGCGGG TACCAACCTGGAAGGCACCGCTGTTGAAATCAAAACCCTGGACGAACTGAAAAACGCCCTGAAA GGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGAAAAAGGCGAAGTCTCGGAAAAAATCATTACCCGTGCTTGCGGCACCCGTCTGGAATAC ACCGGCATTAAATCGGATGGCAGCGGCAAAGCGAAGGAAGTTCTGAAAAACTTTACCCTGGAAG GCAAAGTCGCAAATGATAAGACCACCCTGGTGGTGAAAGAAGGCACCGTTACGCTGAGCAAAAA CATTAGTAAGTCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGCCACCAAA AAGACGGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATTCCAAAAAGACCA AAGATCTGGTCTTCACGAAAGAATGCACCATCACGGTGCAGCAATATGACAGCAACGGTACCAA ACTGGAAGGCTCTGCGGTGGAAATCACGAAACTGGATGAAATCAAAAATGCACTGAAAGGTCTC GAGCACCACCACCACCACCAC SEQIDNO:71 S2D1-S1D1_aa:HeterodimerfusionproteinofOspAserotypes2and1bothwithdisulfidebondtype 1,N-terminalCSSforadditionoflipids,LN1linkersequence,aa164-174ofOspAserotype1 replacedbynon-hLFA-1-likesequenceNFTLEGKVAND FNEKGELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKCGTVTLSKEIAK SGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEI KTLDELCNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEVSEKIITRADGTRLEYTGIKSDGSGKAKEV LKNFTLEGKVANDKTTLVVKCGTVTLSKNISKSGEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSK KTKDLVFTKENTITVQQYDSNGTKLEGSAVEITKLDEICNALK SEQIDNO:72 Lip-S2D1-S1D1_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotypes2and1bothwithdisulfidebondtype1,E.coliIpplipidationsignal,N-terminalCSSfor additionoflipids,LN1linkersequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-like sequenceNFTLEGKVAND ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAAAACGGCACCAAACT GGAATATACGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCC TGGAAGGCAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAATGCGGCACCGTTACGCTGTC AAAAGAAATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCG ACCAAGAAAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAATAGCAAGA AAACCACGCAGCTGGTCTTCACCAAACAAGATACGATCACCGTGCAGAAATACGACAGTGCGGG TACCAACCTGGAAGGCACGGCTGTTGAAATCAAAACCCTGGACGAACTGTGTAACGCCCTGAAA GGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGAAAAAGGCGAAGTCAGCGAAAAAATCATTACCCGCGCAGACGGCACCCGCCTGGAATAC ACCGGCATCAAATCGGACGGCAGCGGCAAAGCGAAAGAAGTTCTGAAAAACTTTACCCTGGAAG GCAAAGTCGCAAATGATAAAACCACCCTGGTGGTGAAATGCGGCACCGTTACGCTGAGCAAAAA CATTAGTAAATCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGCCACCAAG AAAACCGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATAGCAAGAAAACCA AAGATCTGGTCTTCACGAAAGAAAACACCATCACGGTGCAGCAATATGACAGCAATGGTACCAAA CTGGAAGGCTCCGCTGTGGAAATCACGAAACTGGATGAAATCTGTAATGCACTGAAA SEQIDNO:73 Lip-S2D1-S1D1_His_aa:HeterodimerfusionproteinofOspAserotypes2and1bothwithdisulfide bondtype1,N-terminalCSSforadditionoflipids,LN1linkersequence,aa164-174ofOspAserotype 1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND,N-terminallipidation,C-terminalHistag (GLEHHHHHH) LipCSSFNEKGELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKCGTVTL SKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLE GTAVEIKTLDELCNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEVSEKIITRADGTRLEYTGIKSDGS GKAKEVLKNFTLEGKVANDKTTLVVKCGTVTLSKNISKSGEVSVELNDTDSSAATKKTAAWNSGTSTL TITVNSKKTKDLVFTKENTITVQQYDSNGTKLEGSAVEITKLDEICNALKGLEHHHHHH SEQIDNO:74 Lip-S2D1-S1D1_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotypes2and1 bothwithdisulfidebondtype1,E.coliIpplipidationsignal,N-terminalCSSforadditionof lipids,LN1linkersequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequence NFTLEGKVAND,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAAAACGGCACCAAACT GGAATATACGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCC TGGAAGGCAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAATGCGGCACCGTTACGCTGTC AAAAGAAATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCG ACCAAGAAAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAATAGCAAGA AAACCACGCAGCTGGTCTTCACCAAACAAGATACGATCACCGTGCAGAAATACGACAGTGCGGG TACCAACCTGGAAGGCACGGCTGTTGAAATCAAAACCCTGGACGAACTGTGTAACGCCCTGAAA GGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGAAAAAGGCGAAGTCAGCGAAAAAATCATTACCCGCGCAGACGGCACCCGCCTGGAATAC ACCGGCATCAAATCGGACGGCAGCGGCAAAGCGAAAGAAGTTCTGAAAAACTTTACCCTGGAAG GCAAAGTCGCAAATGATAAAACCACCCIGGTGGTGAAATGCGGCACCGTTACGCTGAGCAAAAA CATTAGTAAATCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGCCACCAAG AAAACCGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATAGCAAGAAAACCA AAGATCTGGTCTTCACGAAAGAAAACACCATCACGGTGCAGCAATATGACAGCAATGGTACCAAA CTGGAAGGCTCCGCTGTGGAAATCACGAAACTGGATGAAATCTGTAATGCACTGAAAGGTCTCG AGCACCACCACCACCACCAC SEQIDNO:75 S4D4-S3D4_aa:HeterodimerfusionproteinofOspAserotypes4and3bothwithdisulfidebondtype 4,N-terminalCSSforadditionoflipids,LN1linkersequence FNAKGELSEKTILRACGTRLEYTEIKSDGTGKAKEVLKDFALEGTLAADKTTLKVTEGTVVLSKHIPNS GEITVELNDSNSTQATKKTGKWDSNTSTLTISVNSKKTKNIVFTKECTITVQKYDSAGTNLEGNAVEIK TLDELKNALKGTSDKNNGSGSKEKNKDGKYSFNDKGKLSEKVVTRACGTRLEYTEIKNDGSGKAKEV LKGFALEGTLTDGGETKLTVTEGTVTLSKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNS QKPKQLVFTKECTITVQNYNRAGNALEGSPAEIKDLAELKAALK SEQIDNO:76 Lip-S4D4-S3D4_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotypes4and3bothwithdisulfidebondtype4,E.coliIpplipidationsignal,N-terminalCSSfor additionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTICAACGCTAAAGGTGAACTGTCGGAAAAAACCATCCTGCGCGCCTGTGGCACCCGCCT GGAATACACGGAAATCAAGTCGGACGGCACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCT CTGGAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTTCTG AGCAAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGC GACCAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAA AAGACCAAAAATATTGTGTTCACGAAGGAATGCACGATCACCGTTCAAAAATATGATTCCGCAGG TACCAACCTGGAAGGCAACGCTGTGGAAATCAAAACCCTGGACGAACTGAAAAACGCCCTGAAG GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT TAACGATAAGGGCAAACTGTCAGAAAAAGTGGTCACCCGCGCTIGTGGCACCCGCCTGGAATAC ACCGAAATCAAAAACGACGGCTCGGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCCCTGGAAG GTACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGGAAGGCACCGTTACGCTGTCTAA AAACATTAGCAAGTCTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCCGGCTGAC AAAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCGCAGAAAC CGAAGCAACTGGTCTTCACCAAAGAATGCACGATCACCGTGCAGAACTATAATCGTGCCGGTAAT GCTCTGGAAGGCTCCCCGGCTGAAATCAAGGACCTGGCGGAACTGAAGGCGGCACTGAAA SEQIDNO:77 Lip-S4D4-S3D4_His_aa:HeterodimerfusionproteinofOspAserotypes4and3bothwithdisulfide bondtype4,N-terminalCSSforadditionoflipids,LN1linkersequence,N-terminallipidation,C- terminalHistag(GLEHHHHHH) LipCSSFNAKGELSEKTILRACGTRLEYTEIKSDGTGKAKEVLKDFALEGTLAADKTTLKVTEGTVVLSK HIPNSGEITVELNDSNSTQATKKTGKWDSNTSTLTISVNSKKTKNIVFTKECTITVQKYDSAGTNLEGN AVEIKTLDELKNALKGTSDKNNGSGSKEKNKDGKYSFNDKGKLSEKVVTRACGTRLEYTEIKNDGSG KAKEVLKGFALEGTLTDGGETKLTVTEGTVTLSKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLT ISKNSQKPKQLVFTKECTITVQNYNRAGNALEGSPAEIKDLAELKAALKGLEHHHHHH SEQIDNO:78 Lip-S4D4-S3D4_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotypes4and3 bothwithdisulfidebondtype4,E.coliIpplipidationsignal,N-terminalCSSforadditionof lipids,LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGCTAAAGGTGAACTGTCGGAAAAAACCATCCTGCGCGCCTGTGGCACCCGCCT GGAATACACGGAAATCAAGTCGGACGGCACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCT CTGGAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTTCTG AGCAAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGC GACCAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAA AAGACCAAAAATATTGTGTTCACGAAGGAATGCACGATCACCGTTCAAAAATATGATTCCGCAGG TACCAACCTGGAAGGCAACGCTGTGGAAATCAAAACCCTGGACGAACTGAAAAACGCCCTGAAG GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT TAACGATAAGGGCAAACTGTCAGAAAAAGTGGTCACCCGCGCTTGTGGCACCCGCCTGGAATAC ACCGAAATCAAAAACGACGGCTCGGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCCCTGGAAG GTACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGGAAGGCACCGTTACGCTGTCTAA AAACATTAGCAAGTCTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCCGGCTGAC AAAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCGCAGAAAC CGAAGCAACTGGTCTTCACCAAAGAATGCACGATCACCGTGCAGAACTATAATCGTGCCGGTAAT GCTCTGGAAGGCTCCCCGGCTGAAATCAAGGACCTGGCGGAACTGAAGGCGGCACTGAAAGGT CTCGAGCACCACCACCACCACCAC SEQIDNO:79 S4D1-S3D1_aa:HeterodimerfusionproteinofOspAserotypes4and3bothwithdisulfidebondtype 1,LN1linkersequence FNAKGELSEKTILRANGTRLEYTEIKSDGTGKAKEVLKDFALEGTLAADKTTLKVTCGTVVLSKHIPNS GEITVELNDSNSTQATKKTGKWDSNTSTLTISVNSKKTKNIVFTKEDTITVQKYDSAGTNLEGNAVEIK TLDELCNALKGTSDKNNGSGSKEKNKDGKYSFNDKGKLSEKVVTRANGTRLEYTEIKNDGSGKAKEV LKGFALEGTLTDGGETKLTVTCGTVTLSKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNS QKPKQLVFTKENTITVQNYNRAGNALEGSPAEIKDLAELCAALK SEQIDNO:80 Lip-S4D1-S3D1_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotypes4and3bothwithdisulfidebondtype1,E.coliIpplipidationsignal,N-terminalCSSfor additionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAATGCTAAGGGCGAACTGAGCGAAAAAACGATCCTGCGTGCGAATGGCACCCGTCT GGAATACACCGAAATCAAATCCGATGGTACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCT CTGGAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGTTCTG AGCAAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGC AACCAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAAA AGACCAAAAATATTGTGTTCACGAAGGAAGATACGATCACCGTTCAAAAATATGACTCCGCGGGC ACCAACCTGGAAGGCAATGCCGTCGAAATCAAAACCCTGGATGAACTGTGTAACGCCCTGAAGG GTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATTT AACGATAAGGGCAAACTGTCGGAAAAAGTGGTCACCCGCGCAAATGGCACCCGCCTGGAATACA CGGAAATCAAAAACGATGGTAGCGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCCCTGGAAGG TACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGTGCGGCACCGTTACGCTGTCTAAA AACATTAGCAAGTCTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCCGGCTGACA AAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCGCAGAAACC GAAGCAACTGGTCTTCACCAAAGAAAACACGATCACCGTGCAGAACTATAATCGTGCCGGTAATG CTCTGGAAGGCTCACCGGCTGAAATCAAGGACCTGGCTGAACTGTGTGCGGCACTGAAA SEQIDNO:81 Lip-S4D1-S3D1_His_aa:HeterodimerfusionproteinofOspAserotypes4and3bothwithdisulfide bondtype1,N-terminalCSSforadditionoflipids,LN1linkersequence,N-terminallipidation,C- terminalHistag(GLEHHHHHH) LipCSSFNAKGELSEKTILRANGTRLEYTEIKSDGTGKAKEVLKDFALEGTLAADKTTLKVTCGTVVLSK HIPNSGEITVELNDSNSTQATKKTGKWDSNTSTLTISVNSKKTKNIVFTKEDTITVQKYDSAGTNLEGN AVEIKTLDELCNALKGTSDKNNGSGSKEKNKDGKYSFNDKGKLSEKVVTRANGTRLEYTEIKNDGSG KAKEVLKGFALEGTLTDGGETKLTVTCGTVTLSKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLT ISKNSQKPKQLVFTKENTITVQNYNRAGNALEGSPAEIKDLAELCAALKGLEHHHHHH SEQIDNO:82 Lip-S4D1-S3D1_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotypes4and3 bothwithdisulfidebondtype1,E.coliIpplipidationsignal,N-terminalCSSforadditionof lipids,LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAATGCTAAGGGCGAACTGAGCGAAAAAACGATCCTGCGTGCGAATGGCACCCGTCT GGAATACACCGAAATCAAATCCGATGGTACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCT CTGGAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGTICTG AGCAAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGC AACCAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAAA AGACCAAAAATATTGTGTTCACGAAGGAAGATACGATCACCGTTCAAAAATATGACTCCGCGGGC ACCAACCTGGAAGGCAATGCCGTCGAAATCAAAACCCTGGATGAACTGTGTAACGCCCTGAAGG GTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATTT AACGATAAGGGCAAACTGTCGGAAAAAGTGGTCACCCGCGCAAATGGCACCCGCCTGGAATACA CGGAAATCAAAAACGATGGTAGCGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCCCTGGAAGG TACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGTGCGGCACCGTTACGCTGTCTAAA AACATTAGCAAGTCTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCCGGCTGACA AAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCGCAGAAACC GAAGCAACTGGICTTCACCAAAGAAAACACGATCACCGTGCAGAACTATAATCGTGCCGGTAATG CTCTGGAAGGCTCACCGGCTGAAATCAAGGACCTGGCTGAACTGTGTGCGGCACTGAAAGGTCT CGAGCACCACCACCACCACCAC SEQIDNO:83 S6D4-S5D4_aa:HeterodimerfusionproteinofOspAserotypes6and5bothwithdisulfidebondtype 4,LN1linkersequence FNGKGETSEKTIVRACGTRLEYTDIKSDGSGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVVLSKNILK SGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQKTKNLVFTKECTITVQRYDSAGTNLEGKAVEI TTLKELKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEISEKTIVRACGTRLEYTDIKSDKTGKAKEV LKDFTLEGTLAADGKTTLKVTEGTVTLSKNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRT KTKQLVFTKECTITVQNYDSAGTNLEGKAVEITTLKELKNALK SEQIDNO:84 Lip-S6D4-S5D4_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotypes6and5bothwithdisulfidebondtype4,E.coliIpplipidationsignal,N-terminalCSSfor additionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGGCAAAGGTGAAACGAGTGAAAAAACGATTGTTCGCGCCTGTGGCACCCGCCT GGAATACACGGATATCAAGTCGGATGGTTCGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTACG CTGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTT CTGTCAAAAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACGC GTGCTACGAAAAAGACCGGTAAATGGGACAGCAAGACCTCTACGCTGACCATTAGTGTCAACTC CCAGAAAACGAAGAATCTGGTGTTCACCAAAGAATGCACGATCACCGTTCAACGCTATGATAGTG CGGGCACCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGAAGAATGCTCT GAAAGGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACT CATTCAACGAAAAAGGCGAAATCAGTGAAAAAACCATTGTGCGTGCGTGTGGCACCCGTCTGGA ATATACCGACATCAAGAGCGATAAAACGGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGCTGG AAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCGAAGGTACCGTTACGCTGT CCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCTGGATGACACCGATAGCTCTGGCAA CAAAAAGAGCGGTACCTGGGACTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACGAAA ACCAAGCAGCTGGTCTTCACGAAAGAATGCACGATCACCGTGCAAAACTATGATAGCGCAGGTA CCAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGAAGAATGCTCTGAAA SEQIDNO:85 Lip-S6D4-S5D4_His_aa:HeterodimerfusionproteinofOspAserotypes6and5bothwithdisulfide bondtype4,N-terminalCSSforadditionoflipids,LN1linkersequence,N-terminallipidation,C- terminalHistag(GLEHHHHHH) LipCSSFNGKGETSEKTIVRACGTRLEYTDIKSDGSGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVVL SKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQKTKNLVFTKECTITVQRYDSAGTNLE GKAVEITTLKELKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEISEKTIVRACGTRLEYTDIKSDKTG KAKEVLKDFTLEGTLAADGKTTLKVTEGTVTLSKNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTI SKNRTKTKQLVFTKECTITVQNYDSAGTNLEGKAVEITTLKELKNALKGLEHHHHHH SEQIDNO:86 Lip-S6D4-S5D4_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotypes6and5 bothwithdisulfidebondtype4,E.coliIpplipidationsignal,N-terminalCSSforadditionof lipids,LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGGCAAAGGTGAAACGAGTGAAAAAACGATTGTTCGCGCCTGTGGCACCCGCCT GGAATACACGGATATCAAGTCGGATGGTTCGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTACG CTGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTT CTGTCAAAAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACGC GTGCTACGAAAAAGACCGGTAAATGGGACAGCAAGACCTCTACGCTGACCATTAGTGTCAACTC CCAGAAAACGAAGAATCTGGTGTTCACCAAAGAATGCACGATCACCGTTCAACGCTATGATAGTG CGGGCACCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGAAGAATGCTCT GAAAGGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACT CATTCAACGAAAAAGGCGAAATCAGTGAAAAAACCATTGTGCGTGCGTGTGGCACCCGTCTGGA ATATACCGACATCAAGAGCGATAAAACGGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGCTGG AAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCGAAGGTACCGTTACGCTGT CCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCTGGATGACACCGATAGCTCTGGCAA CAAAAAGAGCGGTACCTGGGACTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACGAAA ACCAAGCAGCTGGTCTTCACGAAAGAATGCACGATCACCGTGCAAAACTATGATAGCGCAGGTA CCAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGAAGAATGCTCTGAAAGG TCTCGAGCACCACCACCACCACCAC SEQIDNO:87 S6D1-S5D1_aa:HeterodimerfusionproteinofOspAserotypes6and5bothwithdisulfidebondtype 1,LN1linkersequence FNGKGETSEKTIVRANGTRLEYTDIKSDGSGKAKEVLKDFTLEGTLAADGKTTLKVTCGTVVLSKNILK SGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQKTKNLVFTKEDTITVQRYDSAGTNLEGKAVEI TTLKELCNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEISEKTIVRANGTRLEYTDIKSDKTGKAKEV LKDFTLEGTLAADGKTTLKVTCGTVTLSKNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRT KTKOLVFTKEDTITVQNYDSAGTNLEGKAVEITTLKELCNALK SEQIDNO:88 Lip-S6D1-S5D1_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotypes6and5bothwithdisulfidebondtype1,E.coliIpplipidationsignal,N-terminalCSSfor additionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGGCAAAGGTGAAACGAGCGAAAAGACCATCGTGCGTGCGAACGGTACCCGCC TGGAATATACGGACATTAAATCGGACGGCAGCGGCAAAGCAAAGGAAGTCCTGAAAGATTTTAC GCTGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGT TCTGTCAAAAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACG CGTGCTACGAAAAAGACCGGTAAATGGGATAGCAAGACCTCTACGCTGACCATTAGTGTCAACT CCCAGAAAACGAAGAATCTGGTGTTCACCAAAGAAGATACGATCACCGTTCAACGCTATGACAGT GCGGGCACCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGTGTAATGCTC TGAAAGGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTAC TCATTCAACGAAAAAGGCGAAATCTCAGAAAAAACCATCGTCCGCGCTAACGGCACCCGCCTGG AATACACCGACATCAAATCAGACAAGACCGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGCTG GAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCTGCGGTACCGTTACGCTG TCCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCTGGATGACACCGATAGCTCTGGCA ACAAAAAGAGCGGTACCTGGGATTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACGAAA ACCAAGCAGCTGGTCTTCACGAAAGAAGATACGATCACCGTGCAAAACTATGACAGCGCAGGTA CCAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGTGTAATGCTCTGAAA SEQIDNO:89 Lip-S6D1-S5D1_His_aa:HeterodimerfusionproteinofOspAserotypes6and5bothwithdisulfide bondtype1,LN1linkersequence,N-terminallipidation,C-terminalHistag(GLEHHHHHH) LipCSSFNGKGETSEKTIVRANGTRLEYTDIKSDGSGKAKEVLKDFTLEGTLAADGKTTLKVTCGTVVL SKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQKTKNLVFTKEDTITVQRYDSAGTNLE GKAVEITTLKELCNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEISEKTIVRANGTRLEYTDIKSDKT GKAKEVLKDFTLEGTLAADGKTTLKVTCGTVTLSKNISKSGEITVALDDTDSSGNKKSGTWDSGTSTL TISKNRTKTKQLVFTKEDTITVQNYDSAGTNLEGKAVEITTLKELCNALKGLEHHHHHH SEQIDNO:90 Lip-S6D1-S5D1_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotypes6and5 bothwithdisulfidebondtype1,E.coliIpplipidationsignal,N-terminalCSSforadditionof lipids,LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGGCAAAGGTGAAACGAGCGAAAAGACCATCGTGCGTGCGAACGGTACCCGCC TGGAATATACGGACATTAAATCGGACGGCAGCGGCAAAGCAAAGGAAGTCCTGAAAGATTTTAC GCTGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGT TCTGTCAAAAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACG CGTGCTACGAAAAAGACCGGTAAATGGGATAGCAAGACCTCTACGCTGACCATTAGTGTCAACT CCCAGAAAACGAAGAATCTGGTGTTCACCAAAGAAGATACGATCACCGTTCAACGCTATGACAGT GCGGGCACCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGTGTAATGCTC TGAAAGGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTAC TCATTCAACGAAAAAGGCGAAATCTCAGAAAAAACCATCGTCCGCGCTAACGGCACCCGCCIGG AATACACCGACATCAAATCAGACAAGACCGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGCTG GAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCTGCGGTACCGTTACGCTG TCCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCTGGATGACACCGATAGCTCTGGCA ACAAAAAGAGCGGTACCTGGGATTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACGAAA ACCAAGCAGCTGGTCTTCACGAAAGAAGATACGATCACCGTGCAAAACTATGACAGCGCAGGTA CCAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGTGTAATGCTCTGAAAGGT CTCGAGCACCACCACCACCACCAC SEQIDNO:91 S1D4-S2D1_aa:HeterodimerfusionproteinofOspAserotype1withdisulfidebondtype4andOspA serotype2withdisulfidebondtype1,LN1linkersequence,aa164-174ofOspAserotype1replaced bynon-hLFA-1-likesequenceNFTLEGKVAND FNEKGEVSEKIITRACGTRLEYTGIKSDGSGKAKEVLKNFTLEGKVANDKTTLVVKEGTVTLSKNISKS GEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSKKTKDLVFTKECTITVQQYDSNGTKLEGSAVEIT KLDEIKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGELSAKTMTRENGTKLEYTEMKSDGTGKAKE VLKNFTLEGKVANDKVTLEVKCGTVTLSKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNS KKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDELCNALK SEQIDNO:92 Lip-S1D4-S2D1_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotype1withdisulfidebondtype4andOspAserotype2withdisulfidebondtype1,E.coliIpp lipidationsignal,N-terminalCSSforadditionoflipids,LN1linkersequence,aa164-174ofOspA serotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAGTCTCGGAAAAAATCATTACCCGTGCTTGCGGCACCCGTCT GGAATACACCGGCATTAAATCGGATGGCAGCGGCAAAGCGAAGGAAGTTCTGAAAAACTTTACC CTGGAAGGCAAAGTCGCAAATGATAAGACCACCCTGGTGGTGAAAGAAGGCACCGTTACGCTGA GCAAAAACATTAGTAAGTCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGC CACCAAAAAGACGGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATTCCAAA AAGACCAAAGATCTGGTCTTCACGAAAGAATGCACCATCACGGTGCAGCAATATGACAGCAACG GTACCAAACTGGAAGGCTCTGCGGTGGAAATCACGAAACTGGATGAAATCAAAAATGCTCTGAAA GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAAAACGGCACCAAACTGGAATAT ACGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCCTGGAAG GCAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAATGCGGCACCGTTACGCTGTCAAAAGA AATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCGACCAAG AAAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAATAGCAAGAAAACCA CGCAGCTGGTCTTCACCAAACAAGATACGATCACCGTGCAGAAATACGACAGTGCGGGTACCAA CCTGGAAGGCACGGCTGTTGAAATCAAAACCCTGGACGAACTGTGTAACGCCCTGAAA SEQIDNO:93 Lip-S1D4-S2D1_His_aa:HeterodimerfusionproteinofOspAserotype1withdisulfidebondtype4 andOspAserotype2withdisulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linker sequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND, N-terminallipidation,C-terminalHistag(GLEHHHHHH) LipCSSFNEKGEVSEKIITRACGTRLEYTGIKSDGSGKAKEVLKNFTLEGKVANDKTTLVVKEGTVTLSK NISKSGEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSKKTKDLVFTKECTITVQQYDSNGTKLEG SAVEITKLDEIKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGELSAKTMTRENGTKLEYTEMKSDGT GKAKEVLKNFTLEGKVANDKVTLEVKCGTVTLSKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTL TISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDELCNALKGLEHHHHHH SEQIDNO:94 Lip-S1D4-S2D1_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotype1with disulfidebondtype4andOspAserotype2withdisulfidebondtype1,E.coliIpplipidation signal,N-terminalCSSforadditionoflipids,LN1linkersequence,aa164-174ofOspAserotype 1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAGTCTCGGAAAAAATCATTACCCGTGCTTGCGGCACCCGTCT GGAATACACCGGCATTAAATCGGATGGCAGCGGCAAAGCGAAGGAAGTTCTGAAAAACTTTACC CTGGAAGGCAAAGTCGCAAATGATAAGACCACCCTGGTGGTGAAAGAAGGCACCGTTACGCTGA GCAAAAACATTAGTAAGTCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGC CACCAAAAAGACGGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATTCCAAA AAGACCAAAGATCTGGTCTTCACGAAAGAATGCACCATCACGGTGCAGCAATATGACAGCAACG GTACCAAACTGGAAGGCTCTGCGGTGGAAATCACGAAACTGGATGAAATCAAAAATGCTCTGAAA GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAAAACGGCACCAAACTGGAATAT ACGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCCTGGAAG GCAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAATGCGGCACCGTTACGCTGTCAAAAGA AATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCGACCAAG AAAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAATAGCAAGAAAACCA CGCAGCTGGTCTTCACCAAACAAGATACGATCACCGTGCAGAAATACGACAGTGCGGGTACCAA CCTGGAAGGCACGGCTGTTGAAATCAAAACCCTGGACGAACTGTGTAACGCCCTGAAAGGCCTC GAGCACCACCACCACCACCAC SEQIDNO:95 S1D1-S2D4_aa:HeterodimerfusionproteinofOspAserotype1withdisulfidebondtype1andOspA serotype2withdisulfidebondtype4,LN1linkersequence,aa164-174ofOspAserotype1replaced bynon-hLFA-1-likesequenceNFTLEGKVAND FNEKGEVSEKIITRADGTRLEYTGIKSDGSGKAKEVLKNFTLEGKVANDKTTLVVKCGTVTLSKNISKS GEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSKKTKDLVFTKENTITVQQYDSNGTKLEGSAVEIT KLDEICNALKGTSDKNNGSGSKEKNKDGKYSFNEKGELSAKTMTRECGTKLEYTEMKSDGTGKAKE VLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVTVALNDINTTQATKKTGAWDSKTSTLTISVNS KKTTQLVFTKQCTITVQKYDSAGTNLEGTAVEIKTLDELKNALK SEQIDNO:96 Lip-S1D1-S2D4_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotype1withdisulfidebondtype1andOspAserotype2withdisulfidebondtype4,E.coliIpp lipidationsignal,N-terminalCSSforadditionoflipids,LN1linkersequence,aa164-174ofOspA serotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAGTCAGCGAAAAAATCATTACCCGCGCAGACGGCACCCGCCT GGAATACACCGGCATCAAATCGGACGGCAGCGGCAAAGCGAAAGAAGTTCTGAAAAACTTTACC CTGGAAGGCAAAGTCGCAAATGATAAAACCACCCTGGTGGTGAAATGCGGCACCGTTACGCTGA GCAAAAACATTAGTAAATCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGC CACCAAGAAAACCGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATAGCAAG AAAACCAAAGATCTGGTCTTCACGAAAGAAAACACCATCACGGTGCAGCAATATGACAGCAATGG TACCAAACTGGAAGGCTCCGCTGTGGAAATCACGAAACTGGATGAAATCTGTAATGCTCTGAAAG GTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATTC AACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAATGCGGCACCAAACTGGAATATA CGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCCTGGAAGG CAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAAGAAGGCACCGTTACGCTGTCAAAAGAA ATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCGACCAAGA AAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAACAGCAAGAAAACCAC GCAGCTGGTCTTCACCAAACAATGTACGATCACCGTGCAGAAATACGATAGTGCGGGTACCAAC CTGGAAGGCACCGCTGTTGAAATCAAAACCCTGGACGAACTGAAAAACGCCCTGAAA SEQIDNO:97 Lip-S1D1-S2D4_His_aa:HeterodimerfusionproteinofOspAserotype1withdisulfidebondtype1 andOspAserotype2withdisulfidebondtype4,N-terminalCSSforadditionoflipids,LN1linker sequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND, N-terminallipidation,C-terminalHistag(GLEHHHHHH) LipCSSFNEKGEVSEKIITRADGTRLEYTGIKSDGSGKAKEVLKNFTLEGKVANDKTTLVVKCGTVTLSK NISKSGEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSKKTKDLVFTKENTITVQQYDSNGTKLEG SAVEITKLDEICNALKGTSDKNNGSGSKEKNKDGKYSFNEKGELSAKTMTRECGTKLEYTEMKSDGT GKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTL TISVNSKKTTQLVFTKQCTITVQKYDSAGTNLEGTAVEIKTLDELKNALKGLEHHHHHH SEQIDNO:98 Lip-S1D1-S2D4_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotype1with disulfidebondtype1andOspAserotype2withdisulfidebondtype4,E.coliIpplipidationsignal, N-terminalCSSforadditionoflipids,LN1linkersequence,aa164-174ofOspAserotype1replaced bynon-hLFA-1-likesequenceNFTLEGKVAND,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAGTCAGCGAAAAAATCATTACCCGCGCAGACGGCACCCGCCT GGAATACACCGGCATCAAATCGGACGGCAGCGGCAAAGCGAAAGAAGTICTGAAAAACTTTACC CTGGAAGGCAAAGTCGCAAATGATAAAACCACCCTGGTGGTGAAATGCGGCACCGTTACGCTGA GCAAAAACATTAGTAAATCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGC CACCAAGAAAACCGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATAGCAAG AAAACCAAAGATCTGGTCTTCACGAAAGAAAACACCATCACGGTGCAGCAATATGACAGCAATGG TACCAAACTGGAAGGCTCCGCTGTGGAAATCACGAAACTGGATGAAATCTGTAATGCTCTGAAAG GTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATTC AACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAATGCGGCACCAAACTGGAATATA CGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCCTGGAAGG CAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAAGAAGGCACCGTTACGCTGTCAAAAGAA ATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCGACCAAGA AAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAACAGCAAGAAAACCAC GCAGCTGGTCTTCACCAAACAATGTACGATCACCGTGCAGAAATACGATAGTGCGGGTACCAAC CTGGAAGGCACCGCTGTTGAAATCAAAACCCTGGACGAACTGAAAAACGCCCTGAAAGGCCTCG AGCACCACCACCACCACCAC SEQIDNO:99 S3D4-S4D1_aa:HeterodimerfusionproteinofOspAserotype3withdisulfidebondtype4andOspA serotype4withdisulfidebondtype1,LN1linkersequence FNEKGKLSEKVVTRACGTRLEYTEIKNDGSGKAKEVLKGFALEGTLTDGGETKLTVTEGTVTLSKNIS KSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNSQKPKQLVFTKECTITVQNYNRAGNALEGSPA EIKDLAELKAALKGTSDKNNGSGSKEKNKDGKYSFNAKGELSEKTILRANGTRLEYTEIKSDGTGKAK EVLKDFALEGTLAADKTTLKVTCGTVVLSKHIPNSGEITVELNDSNSTQATKKTGKWDSNTSTLTISVN SKKTKNIVFTKEDTITVQKYDSAGTNLEGNAVEIKTLDELCNALK SEQIDNO:100 Lip-S3D4-S4D1_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotype3withdisulfidebondtype4andOspAserotype4withdisulfidebondtype1,E.coliIpp lipidationsignal,N-terminalCSSforadditionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCAAACTGTCAGAAAAAGTGGTCACCCGCGCTTGTGGCACCCGCCT GGAATACACCGAAATCAAAAACGACGGCTCGGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCC CTGGAAGGTACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGGAAGGCACCGTTACG CTGTCTAAAAACATTAGCAAGICTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCC GGCTGACAAAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCG CAGAAACCGAAGCAACTGGTCTTCACCAAAGAATGCACGATCACCGTGCAGAACTATAATCGTG CCGGTAATGCTCTGGAAGGCTCCCCGGCTGAAATCAAGGACCTGGCGGAACTGAAGGCGGCAC TGAAAGGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTA CTCATTCAACGCTAAAGGTGAACTGAGCGAAAAAACGATCCTGCGTGCGAATGGCACCCGTCTG GAATACACCGAAATCAAATCCGATGGTACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCTCT GGAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGTTCTGAG CAAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGCAA CCAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAAAA GACCAAAAATATTGTGTTCACGAAGGAAGATACGATCACCGTTCAAAAATATGACTCCGCGGGCA CCAACCTGGAAGGCAATGCCGTCGAAATCAAAACCCTGGATGAACTGTGTAATGCTCTGAAG SEQIDNO:101 Lip-S3D4-S4D1_His_aa:HeterodimerfusionproteinofOspAserotype3withdisulfidebondtype4 andOspAserotype4withdisulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminallipidation,C-terminalHistag(GLEHHHHHH) LipCSSFNEKGKLSEKVVTRACGTRLEYTEIKNDGSGKAKEVLKGFALEGTLTDGGETKLTVTEGTVTL SKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNSQKPKQLVFTKECTITVQNYNRAGNALE GSPAEIKDLAELKAALKGTSDKNNGSGSKEKNKDGKYSFNAKGELSEKTILRANGTRLEYTEIKSDGT GKAKEVLKDFALEGTLAADKTTLKVTCGTVVLSKHIPNSGEITVELNDSNSTQATKKTGKWDSNTSTL TISVNSKKTKNIVFTKEDTITVQKYDSAGTNLEGNAVEIKTLDELCNALKGLEHHHHHH SEQIDNO:102 Lip-S3D4-S4D1_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotype3with disulfidebondtype4andOspAserotype4withdisulfidebondtype1,E.coliIpplipidationsignal, N-terminalCSSforadditionoflipids,LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCAAACTGTCAGAAAAAGTGGTCACCCGCGCTTGTGGCACCCGCCT GGAATACACCGAAATCAAAAACGACGGCTCGGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCC CTGGAAGGTACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGGAAGGCACCGTTACG CTGTCTAAAAACATTAGCAAGTCTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCC GGCTGACAAAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCG CAGAAACCGAAGCAACTGGTCTTCACCAAAGAATGCACGATCACCGTGCAGAACTATAATCGTG CCGGTAATGCTCTGGAAGGCTCCCCGGCTGAAATCAAGGACCTGGCGGAACTGAAGGCGGCAC TGAAAGGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTA CTCATTCAACGCTAAAGGTGAACTGAGCGAAAAAACGATCCTGCGTGCGAATGGCACCCGTCTG GAATACACCGAAATCAAATCCGATGGTACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCTCT GGAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGTTCTGAG CAAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGCAA CCAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAAAA GACCAAAAATATTGTGTTCACGAAGGAAGATACGATCACCGTTCAAAAATATGACTCCGCGGGCA CCAACCTGGAAGGCAATGCCGTCGAAATCAAAACCCTGGATGAACTGTGTAATGCTCTGAAGGG TCTCGAGCACCACCACCACCACCAC SEQIDNO:103 S3D1-S4D4_aa:HeterodimerfusionproteinofOspAserotype3withdisulfidebondtype1andOspA serotype4withdisulfidebondtype1,LN1linkersequence FNEKGKLSEKVVTRANGTRLEYTEIKNDGSGKAKEVLKGFALEGTLTDGGETKLTVTCGTVTLSKNIS KSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNSQKPKQLVFTKENTITVQNYNRAGNALEGSPA EIKDLAELCAALKGTSDKNNGSGSKEKNKDGKYSFNAKGELSEKTILRACGTRLEYTEIKSDGTGKAK EVLKDFALEGTLAADKTTLKVTEGTVVLSKHIPNSGEITVELNDSNSTQATKKTGKWDSNTSTLTISVN SKKTKNIVFTKECTITVQKYDSAGTNLEGNAVEIKTLDELKNALK SEQIDNO:104 Lip-S3D1-S4D4_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotype3withdisulfidebondtype1andOspAserotype4withdisulfidebondtype1,E.coliIpp lipidationsignal,N-terminalCSSforadditionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTICAACGAAAAGGGCAAACTGTCGGAAAAAGIGGTCACCCGCGCAAATGGCACCCGCCT GGAATACACGGAAATCAAAAACGATGGTAGCGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCC CTGGAAGGTACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGTGCGGCACCGTTACG CTGTCTAAAAACATTAGCAAGTCTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCC GGCTGACAAAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCG CAGAAACCGAAGCAACTGGTCTTCACCAAAGAAAACACGATCACCGTGCAGAACTATAATCGTGC CGGTAATGCTCTGGAAGGCTCACCGGCTGAAATCAAGGACCTGGCTGAACTGTGTGCGGCACT GAAAGGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTAC TCATTCAACGCTAAAGGTGAACTGTCGGAAAAAACCATCCTGCGCGCCTGTGGCACCCGCCTGG AATACACGGAAATCAAGTCGGACGGCACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCTCT GGAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTTCTGAG CAAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGCGA CCAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAAAA GACCAAAAATATTGTGTTCACGAAGGAATGCACGATCACCGTTCAAAAATATGATTCCGCAGGTA CCAACCTGGAAGGCAACGCTGTGGAAATCAAAACCCTGGACGAACTGAAAAATGCTCTGAAG SEQIDNO:105 Lip-S3D1-S4D4_His_aa:HeterodimerfusionproteinofOspAserotype3withdisulfidebondtype1 andOspAserotype4withdisulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminallipidation,C-terminalHistag(GLEHHHHHH) LipCSSFNEKGKLSEKVVTRANGTRLEYTEIKNDGSGKAKEVLKGFALEGTLTDGGETKLTVTCGTVTL SKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNSQKPKQLVFTKENTITVQNYNRAGNALE GSPAEIKDLAELCAALKGTSDKNNGSGSKEKNKDGKYSFNAKGELSEKTILRACGTRLEYTEIKSDGT GKAKEVLKDFALEGTLAADKTTLKVTEGTVVLSKHIPNSGEITVELNDSNSTQATKKTGKWDSNTSTL TISVNSKKTKNIVFTKECTITVQKYDSAGTNLEGNAVEIKTLDELKNALKGLEHHHHHH SEQIDNO:106 Lip-S3D1-S4D4_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotype3with disulfidebondtype1andOspAserotype4withdisulfidebondtype1,E.coliIpplipidationsignal, N-terminalCSSforadditionoflipids,LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCAAACTGTCGGAAAAAGTGGTCACCCGCGCAAATGGCACCCGCCT GGAATACACGGAAATCAAAAACGATGGTAGCGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCC CTGGAAGGTACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGTGCGGCACCGTTACG CTGTCTAAAAACATTAGCAAGTCTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCC GGCTGACAAAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCG CAGAAACCGAAGCAACTGGTCTTCACCAAAGAAAACACGATCACCGTGCAGAACTATAATCGTGC CGGTAATGCTCTGGAAGGCTCACCGGCTGAAATCAAGGACCTGGCTGAACTGTGTGCGGCACT GAAAGGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTAC TCATTCAACGCTAAAGGTGAACTGTCGGAAAAAACCATCCTGCGCGCCTGTGGCACCCGCCTGG AATACACGGAAATCAAGTCGGACGGCACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCTCT GGAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTTCTGAG CAAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGCGA CCAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAAAA GACCAAAAATATTGTGTTCACGAAGGAATGCACGATCACCGTTCAAAAATATGATTCCGCAGGTA CCAACCTGGAAGGCAACGCTGTGGAAATCAAAACCCTGGACGAACTGAAAAATGCTCTGAAGGG TCTCGAGCACCACCACCACCACCAC SEQIDNO:107 S5D4-S6D1_aa:HeterodimerfusionproteinofOspAserotype5withdisulfidebondtype4andOspA serotype6withdisulfidebondtype1,LN1linkersequence FNEKGEISEKTIVRACGTRLEYTDIKSDKTGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVTLSKNISKS GEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRTKTKQLVFTKECTITVQNYDSAGTNLEGKAVEITT LKELKNALKGTSDKNNGSGSKEKNKDGKYSFNGKGETSEKTIVRANGTRLEYTDIKSDGSGKAKEVL KDFTLEGTLAADGKTTLKVTCGTVVLSKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQ KTKNLVFTKEDTITVQRYDSAGTNLEGKAVEITTLKELCNALK SEQIDNO:108 Lip-S5D4-S6D1_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotype5withdisulfidebondtype4andOspAserotype6withdisulfidebondtype1,E.coliIpp lipidationsignal,N-terminalCSSforadditionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAATCAGTGAAAAAACCATTGTGCGTGCGTGTGGCACCCGTCT GGAATATACCGACATCAAGAGCGATAAAACGGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGC TGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCGAAGGTACCGTTACGC TGTCCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCIGGATGACACCGATAGCTCTGG CAACAAAAAGAGCGGTACCTGGGACTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACG AAAACCAAGCAGCTGGTCTTCACGAAAGAATGCACGATCACCGTGCAAAACTATGATAGCGCAG GTACCAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGAAGAATGCTCTGAAA GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGGCAAAGGTGAAACGAGCGAAAAGACCATCGTGCGTGCGAACGGTACCCGCCTGGAATA TACGGACATTAAATCGGACGGCAGCGGCAAAGCAAAGGAAGTCCTGAAAGATTTTACGCTGGAA GGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGTTCTGTCA AAAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACGCGTGCTA CGAAAAAGACCGGTAAATGGGATAGCAAGACCTCTACGCTGACCATTAGTGTCAACTCCCAGAA AACGAAGAATCTGGTGTTCACCAAAGAAGATACGATCACCGTTCAACGCTATGACAGTGCGGGC ACCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGTGTAATGCTCTGAAA SEQIDNO:109 Lip-S5D4-S6D1_His_aa:HeterodimerfusionproteinofOspAserotype5withdisulfidebondtype4 andOspAserotype6withdisulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminallipidation,C-terminalHistag(GLEHHHHHH) LipCSSFNEKGEISEKTIVRACGTRLEYTDIKSDKTGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVTLS KNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRTKTKQLVFTKECTITVQNYDSAGTNLEGK AVEITTLKELKNALKGTSDKNNGSGSKEKNKDGKYSFNGKGETSEKTIVRANGTRLEYTDIKSDGSGK AKEVLKDFTLEGTLAADGKTTLKVTCGTVVLSKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTIS VNSQKTKNLVFTKEDTITVQRYDSAGTNLEGKAVEITTLKELCNALKGLEHHHHHH SEQIDNO:110 Lip-S5D4-S6D1_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotype5with disulfidebondtype4andOspAserotype6withdisulfidebondtype1,E.coliIpplipidationsignal, N-terminalCSSforadditionoflipids,LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTICAACGAAAAGGGCGAAATCAGTGAAAAAACCATTGTGCGTGCGTGTGGCACCCGTCT GGAATATACCGACATCAAGAGCGATAAAACGGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGC TGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCGAAGGTACCGTTACGC TGTCCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCTGGATGACACCGATAGCTCTGG CAACAAAAAGAGCGGTACCTGGGACTCAGGCACCTCGACGCTGACCATTICTAAAAATCGTACG AAAACCAAGCAGCTGGTCTTCACGAAAGAATGCACGATCACCGTGCAAAACTATGATAGCGCAG GTACCAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGAAGAATGCTCTGAAA GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGGCAAAGGTGAAACGAGCGAAAAGACCATCGTGCGTGCGAACGGTACCCGCCTGGAATA TACGGACATTAAATCGGACGGCAGCGGCAAAGCAAAGGAAGTCCTGAAAGATTTTACGCTGGAA GGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGTTCTGTCA AAAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACGCGTGCTA CGAAAAAGACCGGTAAATGGGATAGCAAGACCTCTACGCTGACCATTAGTGTCAACTCCCAGAA AACGAAGAATCTGGTGTTCACCAAAGAAGATACGATCACCGTTCAACGCTATGACAGTGCGGGC ACCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGTGTAATGCTCTGAAAG GTCTCGAGCACCACCACCACCACCAC SEQIDNO:111 S5D1-S6D4_aa:HeterodimerfusionproteinofOspAserotype5withdisulfidebondtype1andOspA serotype6withdisulfidebondtype4,LN1linkersequence FNEKGEISEKTIVRANGTRLEYTDIKSDKTGKAKEVLKDFTLEGTLAADGKTTLKVTCGTVTLSKNISKS GEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRTKTKQLVFTKEDTITVQNYDSAGTNLEGKAVEITT LKELCNALKGTSDKNNGSGSKEKNKDGKYSFNGKGETSEKTIVRACGTRLEYTDIKSDGSGKAKEVL KDFTLEGTLAADGKTTLKVTEGTVVLSKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQ KTKNLVFTKECTITVQRYDSAGTNLEGKAVEITTLKELKNALK SEQIDNO:112 Lip-S5D1-S6D4_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotype5withdisulfidebondtype1andOspAserotype6withdisulfidebondtype4,E.coliIpp lipidationsignal,N-terminalCSSforadditionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAATCTCAGAAAAAACCATCGTCCGCGCTAACGGCACCCGCCT GGAATACACCGACATCAAATCAGACAAGACCGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGC TGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCTGCGGTACCGTTACGC TGTCCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCTGGATGACACCGATAGCTCTGG CAACAAAAAGAGCGGTACCTGGGATTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACG AAAACCAAGCAGCTGGTCTTCACGAAAGAAGATACGATCACCGTGCAAAACTATGACAGCGCAG GTACCAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGTGTAATGCTCTGAAA GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGGCAAAGGTGAAACGAGTGAAAAAACGATTGTTCGCGCCTGTGGCACCCGCCTGGAATAC ACGGATATCAAGTCGGATGGTTCGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTACGCTGGAAG GTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTTCTGTCAA AAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACGCGTGCTAC GAAAAAGACCGGTAAATGGGACAGCAAGACCTCTACGCTGACCATTAGTGTCAACTCCCAGAAA ACGAAGAATCTGGTGTTCACCAAAGAATGCACGATCACCGTTCAACGCTATGATAGTGCGGGCA CCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGAAGAATGCTCTGAAA SEQIDNO:113 Lip-S5D1-S6D4_His_aa:HeterodimerfusionproteinofOspAserotype5withdisulfidebondtype1 andOspAserotype6withdisulfidebondtype4,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminallipidation,C-terminalHistag(GLEHHHHHH) LipCSSFNEKGEISEKTIVRANGTRLEYTDIKSDKTGKAKEVLKDFTLEGTLAADGKTTLKVTCGTVTLS KNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRTKTKQLVFTKEDTITVQNYDSAGTNLEGK AVEITTLKELCNALKGTSDKNNGSGSKEKNKDGKYSFNGKGETSEKTIVRACGTRLEYTDIKSDGSGK AKEVLKDFTLEGTLAADGKTTLKVTEGTVVLSKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTIS VNSQKTKNLVFTKECTITVQRYDSAGTNLEGKAVEITTLKELKNALKGLEHHHHHH SEQIDNO:114 Lip-S5D1-S6D4_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotype5with disulfidebondtype1andOspAserotype6withdisulfidebondtype4,E.coliIpplipidationsignal, N-terminalCSSforadditionoflipids,LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAGGGCGAAATCTCAGAAAAAACCATCGTCCGCGCTAACGGCACCCGCCT GGAATACACCGACATCAAATCAGACAAGACCGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGC TGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCTGCGGTACCGTTACGC TGTCCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCTGGATGACACCGATAGCTCTGG CAACAAAAAGAGCGGTACCTGGGATTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACG AAAACCAAGCAGCTGGTCTTCACGAAAGAAGATACGATCACCGTGCAAAACTATGACAGCGCAG GTACCAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGTGTAATGCTCTGAAA GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGGCAAAGGTGAAACGAGTGAAAAAACGATTGTTCGCGCCTGTGGCACCCGCCTGGAATAC ACGGATATCAAGTCGGATGGTTCGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTACGCTGGAAG GTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTTCTGTCAA AAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACGCGTGCTAC GAAAAAGACCGGTAAATGGGACAGCAAGACCTCTACGCTGACCATTAGTGTCAACTCCCAGAAA ACGAAGAATCTGGTGTTCACCAAAGAATGCACGATCACCGTTCAACGCTATGATAGTGCGGGCA CCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGAAGAATGCTCTGAAAGG TCTCGAGCACCACCACCACCACCAC SEQIDNO:115 S2D4-S1D1_aa:HeterodimerfusionproteinofOspAserotype2withdisulfidebondtype4andOspA serotype1withdisulfidebondtype1,LN1linkersequence,aa164-174ofOspAserotype1replaced bynon-hLFA-1-likesequenceNFTLEGKVAND FNEKGELSAKTMTRECGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAK SGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQCTITVQKYDSAGTNLEGTAVEI KTLDELKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEVSEKIITRADGTRLEYTGIKSDGSGKAKEV LKNFTLEGKVANDKTTLVVKCGTVTLSKNISKSGEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSK KTKDLVFTKENTITVQQYDSNGTKLEGSAVEITKLDEICNALK SEQIDNO:116 Lip-S2D4-S1D1_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotype2withdisulfidebondtype4andOspAserotype1withdisulfidebondtype1,E.coliIpp lipidationsignal,N-terminalCSSforadditionoflipids,LN1linkersequence,aa164-174ofOspA serotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAATGCGGCACCAAACT GGAATATACGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCC TGGAAGGCAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAAGAAGGCACCGTTACGCTGTC AAAAGAAATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCG ACCAAGAAAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAACAGCAAGA AAACCACGCAGCTGGTCTTCACCAAACAATGTACGATCACCGTGCAGAAATACGATAGTGCGGG TACCAACCTGGAAGGCACCGCTGTTGAAATCAAAACCCTGGACGAACTGAAAAACGCCCTGAAA GGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGAAAAAGGCGAAGTCAGCGAAAAAATCATTACCCGCGCAGACGGCACCCGCCTGGAATAC ACCGGCATCAAATCGGACGGCAGCGGCAAAGCGAAAGAAGTTCTGAAAAACTTTACCCTGGAAG GCAAAGTCGCAAATGATAAAACCACCCTGGTGGTGAAATGCGGCACCGTTACGCTGAGCAAAAA CATTAGTAAATCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGCCACCAAG AAAACCGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATAGCAAGAAAACCA AAGATCTGGTCTTCACGAAAGAAAACACCATCACGGTGCAGCAATATGACAGCAATGGTACCAAA CTGGAAGGCTCCGCTGTGGAAATCACGAAACTGGATGAAATCTGTAATGCACTGAAA SEQIDNO:117 Lip-S2D4-S1D1_His_aa:HeterodimerfusionproteinofOspAserotype2withdisulfidebondtype4 andOspAserotype1withdisulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linker sequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND, N-terminallipidation,C-terminalHistag(GLEHHHHHH) LipCSSFNEKGELSAKTMTRECGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTL SKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQCTITVQKYDSAGTNLE GTAVEIKTLDELKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEVSEKIITRADGTRLEYTGIKSDGS GKAKEVLKNFTLEGKVANDKTTLVVKCGTVTLSKNISKSGEVSVELNDTDSSAATKKTAAWNSGTSTL TITVNSKKTKDLVFTKENTITVQQYDSNGTKLEGSAVEITKLDEICNALKGLEHHHHHH SEQIDNO:118 Lip-S2D4-S1D1_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotype2with disulfidebondtype4andOspAserotype1withdisulfidebondtype1,E.coliIpplipidationsignal, N-terminalCSSforadditionoflipids,LN1linkersequence,aa164-174ofOspAserotype1replaced bynon-hLFA-1-likesequenceNFTLEGKVAND,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAATGCGGCACCAAACT GGAATATACGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCC TGGAAGGCAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAAGAAGGCACCGTTACGCTGTC AAAAGAAATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCG ACCAAGAAAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAACAGCAAGA AAACCACGCAGCTGGTCTTCACCAAACAATGTACGATCACCGTGCAGAAATACGATAGTGCGGG TACCAACCTGGAAGGCACCGCTGTTGAAATCAAAACCCTGGACGAACTGAAAAACGCCCTGAAA GGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGAAAAAGGCGAAGTCAGCGAAAAAATCATTACCCGCGCAGACGGCACCCGCCTGGAATAC ACCGGCATCAAATCGGACGGCAGCGGCAAAGCGAAAGAAGTTCTGAAAAACTTTACCCTGGAAG GCAAAGTCGCAAATGATAAAACCACCCTGGTGGTGAAATGCGGCACCGTTACGCTGAGCAAAAA CATTAGTAAATCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGCCACCAAG AAAACCGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATAGCAAGAAAACCA AAGATCTGGTCTTCACGAAAGAAAACACCATCACGGTGCAGCAATATGACAGCAATGGTACCAAA CTGGAAGGCTCCGCTGTGGAAATCACGAAACTGGATGAAATCTGTAATGCACTGAAAGGTCTCG AGCACCACCACCACCACCAC SEQIDNO:119 S2D1-S1D4_aa:HeterodimerfusionproteinofOspAserotype2withdisulfidebondtype1andOspA serotype1withdisulfidebondtype4,LN1linkersequence,aa164-174ofOspAserotype1replaced bynon-hLFA-1-likesequenceNFTLEGKVAND FNEKGELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKCGTVTLSKEIAK SGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEI KTLDELCNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEVSEKIITRACGTRLEYTGIKSDGSGKAKEV LKNFTLEGKVANDKTTLVVKEGTVTLSKNISKSGEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSK KTKDLVFTKECTITVQQYDSNGTKLEGSAVEITKLDEIKNALK SEQIDNO:120 Lip-S2D1-S1D4_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotype2withdisulfidebondtype1andOspAserotype1withdisulfidebondtype4,E.coliIpp lipidationsignal,N-terminalCSSforadditionoflipids,LN1linkersequence,aa164-174ofOspA serotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAAAACGGCACCAAACT GGAATATACGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCC TGGAAGGCAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAATGCGGCACCGTTACGCTGTC AAAAGAAATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCG ACCAAGAAAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAATAGCAAGA AAACCACGCAGCTGGTCTTCACCAAACAAGATACGATCACCGTGCAGAAATACGACAGTGCGGG TACCAACCTGGAAGGCACGGCTGTTGAAATCAAAACCCTGGACGAACTGTGTAACGCCCTGAAA GGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGAAAAAGGCGAAGTCTCGGAAAAAATCATTACCCGTGCTTGCGGCACCCGTCTGGAATAC ACCGGCATTAAATCGGATGGCAGCGGCAAAGCGAAGGAAGTTCTGAAAAACTTTACCCTGGAAG GCAAAGTCGCAAATGATAAGACCACCCTGGTGGTGAAAGAAGGCACCGTTACGCTGAGCAAAAA CATTAGTAAGTCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGCCACCAAA AAGACGGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATTCCAAAAAGACCA AAGATCTGGTCTTCACGAAAGAATGCACCATCACGGTGCAGCAATATGACAGCAACGGTACCAA ACTGGAAGGCTCTGCGGTGGAAATCACGAAACTGGATGAAATCAAAAATGCACTGAAA SEQIDNO:121 Lip-S2D1-S1D4_His_aa:HeterodimerfusionproteinofOspAserotype2withdisulfidebondtype1 andOspAserotype1withdisulfidebondtype4,N-terminalCSSforadditionoflipids,LN1linker sequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND, N-terminallipidation,C-terminalHistag(GLEHHHHHH) LipCSSFNEKGELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKCGTVTL SKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLE GTAVEIKTLDELCNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEVSEKIITRACGTRLEYTGIKSDGS GKAKEVLKNFTLEGKVANDKTTLVVKEGTVTLSKNISKSGEVSVELNDTDSSAATKKTAAWNSGTSTL TITVNSKKTKDLVFTKECTITVQQYDSNGTKLEGSAVEITKLDEIKNALKGLEHHHHHH SEQIDNO:122 Lip-S2D1-S1D4_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotype2with disulfidebondtype1andOspAserotype1withdisulfidebondtype4,E.coliIpplipidationsignal, N-terminalCSSforadditionoflipids,LN1linkersequence,aa164-174ofOspAserotype1replaced bynon-hLFA-1-likesequenceNFTLEGKVAND,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGAAAAAGGCGAACTGTCGGCGAAAACGATGACGCGTGAAAACGGCACCAAACT GGAATATACGGAAATGAAAAGCGATGGCACCGGTAAAGCGAAAGAAGTTCTGAAAAACTTTACCC TGGAAGGCAAAGTCGCCAATGACAAAGTCACCCTGGAAGTGAAATGCGGCACCGTTACGCTGTC AAAAGAAATTGCAAAATCGGGTGAAGTGACCGTTGCTCTGAACGATACGAATACCACGCAAGCG ACCAAGAAAACCGGCGCCTGGGACAGCAAAACCTCTACGCTGACCATTAGTGTTAATAGCAAGA AAACCACGCAGCTGGTCTTCACCAAACAAGATACGATCACCGTGCAGAAATACGACAGTGCGGG TACCAACCTGGAAGGCACGGCTGTTGAAATCAAAACCCTGGACGAACTGTGTAACGCCCTGAAA GGCACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT CAACGAAAAAGGCGAAGTCTCGGAAAAAATCATTACCCGTGCTTGCGGCACCCGTCTGGAATAC ACCGGCATTAAATCGGATGGCAGCGGCAAAGCGAAGGAAGTTCTGAAAAACTTTACCCTGGAAG GCAAAGTCGCAAATGATAAGACCACCCTGGTGGTGAAAGAAGGCACCGTTACGCTGAGCAAAAA CATTAGTAAGTCCGGTGAAGTCTCTGTGGAACTGAATGATACCGACAGCTCTGCGGCCACCAAA AAGACGGCAGCTTGGAACTCAGGCACCTCGACGCTGACCATTACGGTTAATTCCAAAAAGACCA AAGATCTGGTCTTCACGAAAGAATGCACCATCACGGTGCAGCAATATGACAGCAACGGTACCAA ACTGGAAGGCTCTGCGGTGGAAATCACGAAACTGGATGAAATCAAAAATGCACTGAAAGGTCTC GAGCACCACCACCACCACCAC SEQIDNO:123 S4D4-S3D1_aa:HeterodimerfusionproteinofOspAserotype4withdisulfidebondtype4andOspA serotype3withdisulfidebondtype1,LN1linkersequence FNAKGELSEKTILRACGTRLEYTEIKSDGTGKAKEVLKDFALEGTLAADKTTLKVTEGTVVLSKHIPNS GEITVELNDSNSTQATKKTGKWDSNTSTLTISVNSKKTKNIVFTKECTITVQKYDSAGTNLEGNAVEIK TLDELKNALKGTSDKNNGSGSKEKNKDGKYSFNDKGKLSEKVVTRANGTRLEYTEIKNDGSGKAKEV LKGFALEGTLTDGGETKLTVTCGTVTLSKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNS QKPKQLVFTKENTITVQNYNRAGNALEGSPAEIKDLAELCAALK SEQIDNO:124 Lip-S4D4-S3D1_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotype4withdisulfidebondtype4andOspAserotype3withdisulfidebondtype1,E.coliIpp lipidationsignal,N-terminalCSSforadditionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGCTAAAGGTGAACTGTCGGAAAAAACCATCCTGCGCGCCTGTGGCACCCGCCT GGAATACACGGAAATCAAGTCGGACGGCACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCT CTGGAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTTCTG AGCAAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGC GACCAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAA AAGACCAAAAATATTGTGTTCACGAAGGAATGCACGATCACCGTTCAAAAATATGATTCCGCAGG TACCAACCTGGAAGGCAACGCTGTGGAAATCAAAACCCTGGACGAACTGAAAAACGCCCTGAAG GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT TAACGATAAGGGCAAACTGTCGGAAAAAGTGGTCACCCGCGCAAATGGCACCCGCCTGGAATAC ACGGAAATCAAAAACGATGGTAGCGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCCCTGGAAG GTACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGTGCGGCACCGTTACGCTGTCTAA AAACATTAGCAAGTCTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCCGGCTGAC AAAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCGCAGAAAC CGAAGCAACTGGTCTTCACCAAAGAAAACACGATCACCGTGCAGAACTATAATCGTGCCGGTAAT GCTCTGGAAGGCTCACCGGCTGAAATCAAGGACCTGGCTGAACTGTGTGCGGCACTGAAA SEQIDNO:125 Lip-S4D4-S3D1_His_aa:HeterodimerfusionproteinofOspAserotype4withdisulfidebondtype4 andOspAserotype3withdisulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminalCSSforadditionoflipids,N-terminallipidation,C-terminalHistag (GLEHHHHHH) LipCSSFNAKGELSEKTILRACGTRLEYTEIKSDGTGKAKEVLKDFALEGTLAADKTTLKVTEGTVVLSK HIPNSGEITVELNDSNSTQATKKTGKWDSNTSTLTISVNSKKTKNIVFTKECTITVQKYDSAGTNLEGN AVEIKTLDELKNALKGTSDKNNGSGSKEKNKDGKYSFNDKGKLSEKVVTRANGTRLEYTEIKNDGSG KAKEVLKGFALEGTLTDGGETKLTVTCGTVTLSKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLT ISKNSQKPKQLVFTKENTITVQNYNRAGNALEGSPAEIKDLAELCAALKGLEHHHHHH SEQIDNO:126 Lip-S4D4-S3D1_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotype4with disulfidebondtype4andOspAserotype3withdisulfidebondtype1,E.coliIpplipidationsignal, N-terminalCSSforadditionoflipids,LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGCTAAAGGTGAACTGTCGGAAAAAACCATCCTGCGCGCCTGTGGCACCCGCCT GGAATACACGGAAATCAAGTCGGACGGCACGGGCAAAGCAAAGGAAGTCCTGAAAGATITTGCT CTGGAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTTCTG AGCAAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGC GACCAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAA AAGACCAAAAATATTGTGTTCACGAAGGAATGCACGATCACCGTTCAAAAATATGATTCCGCAGG TACCAACCTGGAAGGCAACGCTGTGGAAATCAAAACCCTGGACGAACTGAAAAACGCCCTGAAG GGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATT TAACGATAAGGGCAAACTGTCGGAAAAAGTGGICACCCGCGCAAATGGCACCCGCCTGGAATAC ACGGAAATCAAAAACGATGGTAGCGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCCCTGGAAG GTACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGTGCGGCACCGTTACGCTGTCTAA AAACATTAGCAAGTCTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCCGGCTGAC AAAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCGCAGAAAC CGAAGCAACTGGTCTTCACCAAAGAAAACACGATCACCGTGCAGAACTATAATCGTGCCGGTAAT GCTCTGGAAGGCTCACCGGCTGAAATCAAGGACCTGGCTGAACTGTGTGCGGCACTGAAAGGT CTCGAGCACCACCACCACCACCAC SEQIDNO:127 S4D1-S3D4_aa:HeterodimerfusionproteinofOspAserotype4withdisulfidebondtype1andOspA serotype3withdisulfidebondtype4,LN1linkersequence FNAKGELSEKTILRANGTRLEYTEIKSDGTGKAKEVLKDFALEGTLAADKTTLKVTCGTVVLSKHIPNS GEITVELNDSNSTQATKKTGKWDSNTSTLTISVNSKKTKNIVFTKEDTITVQKYDSAGTNLEGNAVEIK TLDELCNALKGTSDKNNGSGSKEKNKDGKYSFNDKGKLSEKVVTRACGTRLEYTEIKNDGSGKAKEV LKGFALEGTLTDGGETKLTVTEGTVTLSKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNS QKPKQLVFTKECTITVQNYNRAGNALEGSPAEIKDLAELKAALK SEQIDNO:128 Lip-S4D1-S3D4_nt:HeterodimerfusionproteinofOspAserotype4withdisulfidebondtype1and OspAserotype3withdisulfidebondtype4,E.coliIpplipidationsignal,N-terminalCSSfor additionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAATGCTAAGGGCGAACTGAGCGAAAAAACGATCCTGCGTGCGAATGGCACCCGTCT GGAATACACCGAAATCAAATCCGATGGTACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCT CTGGAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGTICTG AGCAAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGC AACCAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAAA AGACCAAAAATATTGTGTTCACGAAGGAAGATACGATCACCGTTCAAAAATATGACTCCGCGGGC ACCAACCTGGAAGGCAATGCCGTCGAAATCAAAACCCTGGATGAACTGTGTAACGCCCTGAAGG GTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATTT AACGATAAGGGCAAACTGTCAGAAAAAGTGGTCACCCGCGCTTGTGGCACCCGCCTGGAATACA CCGAAATCAAAAACGACGGCTCGGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCCCTGGAAG GTACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGGAAGGCACCGTTACGCTGTCTAA AAACATTAGCAAGTCTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCCGGCTGAC AAAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCGCAGAAAC CGAAGCAACTGGTCTTCACCAAAGAATGCACGATCACCGTGCAGAACTATAATCGTGCCGGTAAT GCTCTGGAAGGCTCCCCGGCTGAAATCAAGGACCTGGCGGAACTGAAGGCGGCACTGAAA SEQIDNO:129 Lip-S4D1-S3D4_His_aa:HeterodimerfusionproteinofOspAserotype4withdisulfidebondtype1 andOspAserotype3withdisulfidebondtype4,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminallipidation,C-terminalHistag(GLEHHHHHH) LipCSSFNAKGELSEKTILRANGTRLEYTEIKSDGTGKAKEVLKDFALEGTLAADKTTLKVTCGTVVLSK HIPNSGEITVELNDSNSTQATKKTGKWDSNTSTLTISVNSKKTKNIVFTKEDTITVQKYDSAGTNLEGN AVEIKTLDELCNALKGTSDKNNGSGSKEKNKDGKYSFNDKGKLSEKVVTRACGTRLEYTEIKNDGSG KAKEVLKGFALEGTLTDGGETKLTVTEGTVTLSKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLT ISKNSQKPKQLVFTKECTITVQNYNRAGNALEGSPAEIKDLAELKAALKGLEHHHHHH SEQIDNO:130 Lip-S4D1-S3D4_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotype4with disulfidebondtype1andOspAserotype3withdisulfidebondtype4,E.coliIpplipidationsignal, N-terminalCSSforadditionoflipids,LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAATGCTAAGGGCGAACTGAGCGAAAAAACGATCCTGCGTGCGAATGGCACCCGTCT GGAATACACCGAAATCAAATCCGATGGTACGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTGCT CTGGAAGGTACCCTGGCGGCCGACAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGTTCTG AGCAAACATATTCCGAACTCTGGTGAAATCACCGTTGAACTGAACGATAGCAATTCTACGCAGGC AACCAAAAAGACGGGCAAATGGGACAGTAATACCTCCACGCTGACCATTTCAGTCAACTCGAAAA AGACCAAAAATATTGTGTTCACGAAGGAAGATACGATCACCGTTCAAAAATATGACTCCGCGGGC ACCAACCTGGAAGGCAATGCCGTCGAAATCAAAACCCTGGATGAACTGTGTAACGCCCTGAAGG GTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACTCATTT AACGATAAGGGCAAACTGTCAGAAAAAGTGGTCACCCGCGCTTGTGGCACCCGCCTGGAATACA CCGAAATCAAAAACGACGGCTCGGGCAAAGCGAAGGAAGTTCTGAAAGGCTTTGCCCTGGAAG GTACCCTGACGGATGGCGGTGAAACCAAACTGACCGTGACGGAAGGCACCGTTACGCTGTCTAA AAACATTAGCAAGTCTGGTGAAATCACGGTCGCACTGAATGATACCGAAACCACGCCGGCTGAC AAAAAGACCGGCGAATGGAAAAGTGACACCTCCACGCTGACCATTTCAAAGAACTCGCAGAAAC CGAAGCAACTGGTCTTCACCAAAGAATGCACGATCACCGTGCAGAACTATAATCGTGCCGGTAAT GCTCTGGAAGGCTCCCCGGCTGAAATCAAGGACCTGGCGGAACTGAAGGCGGCACTGAAAGGT CTCGAGCACCACCACCACCACCAC SEQIDNO:131 S6D4-S5D1_aa:HeterodimerfusionproteinofOspAserotype6withdisulfidebondtype4andOspA serotype5withdisulfidebondtype1,LN1linkersequence FNGKGETSEKTIVRACGTRLEYTDIKSDGSGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVVLSKNILK SGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQKTKNLVFTKECTITVQRYDSAGTNLEGKAVEI TTLKELKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEISEKTIVRANGTRLEYTDIKSDKTGKAKEV LKDFTLEGTLAADGKTTLKVTCGTVTLSKNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRT KTKQLVFTKEDTITVQNYDSAGTNLEGKAVEITTLKELCNALK SEQIDNO:132 Lip-S6D4-S5D1_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotype6withdisulfidebondtype4andOspAserotype5withdisulfidebondtype1,E.coliIpp lipidationsignal,N-terminalCSSforadditionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGGCAAAGGTGAAACGAGTGAAAAAACGATTGTTCGCGCCTGTGGCACCCGCCT GGAATACACGGATATCAAGTCGGATGGTTCGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTACG CTGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTT CTGTCAAAAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACGC GTGCTACGAAAAAGACCGGTAAATGGGACAGCAAGACCTCTACGCTGACCATTAGTGTCAACTC CCAGAAAACGAAGAATCTGGTGTTCACCAAAGAATGCACGATCACCGTTCAACGCTATGATAGTG CGGGCACCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGAAGAATGCTCT GAAAGGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACT CATTCAACGAAAAAGGCGAAATCTCAGAAAAAACCATCGTCCGCGCTAACGGCACCCGCCTGGA ATACACCGACATCAAATCAGACAAGACCGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGCTGG AAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCTGCGGTACCGTTACGCTGT CCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCTGGATGACACCGATAGCTCTGGCAA CAAAAAGAGCGGTACCTGGGATTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACGAAAA CCAAGCAGCTGGTCTTCACGAAAGAAGATACGATCACCGTGCAAAACTATGACAGCGCAGGTAC CAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGTGTAATGCTCTGAAA SEQIDNO:133 Lip-S6D4-S5D1_His_aa:HeterodimerfusionproteinofOspAserotype6withdisulfidebondtype4 andOspAserotype5withdisulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminallipidation,C-terminalHistag(GLEHHHHHH) LipCSSFNGKGETSEKTIVRACGTRLEYTDIKSDGSGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVVL SKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQKTKNLVFTKECTITVQRYDSAGTNLE GKAVEITTLKELKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEISEKTIVRANGTRLEYTDIKSDKTG KAKEVLKDFTLEGTLAADGKTTLKVTCGTVTLSKNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTI SKNRTKTKQLVFTKEDTITVQNYDSAGTNLEGKAVEITTLKELCNALKGLEHHHHHH SEQIDNO:134 Lip-S6D4-S5D1_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotype6with disulfidebondtype4andOspAserotype5withdisulfidebondtype1,E.coliIpplipidationsignal, N-terminalCSSforadditionoflipids,LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGGCAAAGGTGAAACGAGTGAAAAAACGATTGTTCGCGCCTGTGGCACCCGCCT GGAATACACGGATATCAAGTCGGATGGTTCGGGCAAAGCAAAGGAAGTCCTGAAAGATTTTACG CTGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGGAAGGCACCGTGGTT CTGTCAAAAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACGC GTGCTACGAAAAAGACCGGTAAATGGGACAGCAAGACCTCTACGCTGACCATTAGTGTCAACTC CCAGAAAACGAAGAATCTGGTGTTCACCAAAGAATGCACGATCACCGTTCAACGCTATGATAGTG CGGGCACCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGAAGAATGCTCT GAAAGGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTACT CATTCAACGAAAAAGGCGAAATCTCAGAAAAAACCATCGTCCGCGCTAACGGCACCCGCCTGGA ATACACCGACATCAAATCAGACAAGACCGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGCTGG AAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCTGCGGTACCGTTACGCTGT CCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCTGGATGACACCGATAGCTCTGGCAA CAAAAAGAGCGGTACCTGGGATTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACGAAAA CCAAGCAGCTGGTCTTCACGAAAGAAGATACGATCACCGTGCAAAACTATGACAGCGCAGGTAC CAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGTGTAATGCTCTGAAAGGTC TCGAGCACCACCACCACCACCAC SEQIDNO:135 S6D1-S5D4_aa:HeterodimerfusionproteinofOspAserotype6withdisulfidebondtype1andOspA serotype5withdisulfidebondtype4,LN1linkersequence FNGKGETSEKTIVRANGTRLEYTDIKSDGSGKAKEVLKDFTLEGTLAADGKTTLKVTCGTVVLSKNILK SGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQKTKNLVFTKEDTITVQRYDSAGTNLEGKAVEI TTLKELCNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEISEKTIVRACGTRLEYTDIKSDKTGKAKEV LKDFTLEGTLAADGKTTLKVTEGTVTLSKNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRT KTKQLVFTKECTITVQNYDSAGTNLEGKAVEITTLKELKNALK SEQIDNO:136 Lip-S6D1-S5D4_nt:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotype6withdisulfidebondtype1andOspAserotype5withdisulfidebondtype4,E.coliIpp lipidationsignal,N-terminalCSSforadditionoflipids,LN1linkersequence ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGGCAAAGGTGAAACGAGCGAAAAGACCATCGTGCGTGCGAACGGTACCCGCC TGGAATATACGGACATTAAATCGGACGGCAGCGGCAAAGCAAAGGAAGTCCTGAAAGATTTTAC GCTGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGT TCTGTCAAAAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACG CGTGCTACGAAAAAGACCGGTAAATGGGATAGCAAGACCTCTACGCTGACCATTAGTGTCAACT CCCAGAAAACGAAGAATCTGGTGTICACCAAAGAAGATACGATCACCGTTCAACGCTATGACAGT GCGGGCACCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGTGTAATGCTC TGAAAGGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTAC TCATTCAACGAAAAAGGCGAAATCAGTGAAAAAACCATTGTGCGTGCGTGTGGCACCCGTCTGG AATATACCGACATCAAGAGCGATAAAACGGGTAAAGCGAAGGAAGTICTGAAAGATTTTACGCTG GAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCGAAGGTACCGTTACGCTG TCCAAAAACATTAGTAAGTCCGGCGAAATCACGGTCGCCCTGGATGACACCGATAGCTCTGGCA ACAAAAAGAGCGGTACCTGGGACTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACGAA AACCAAGCAGCTGGTCTTCACGAAAGAATGCACGATCACCGTGCAAAACTATGATAGCGCAGGT ACCAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGAAGAATGCTCTGAAA SEQIDNO:137 Lip-S6D1-S5D4_His_aa:HeterodimerfusionproteinofOspAserotype6withdisulfidebondtype1 andOspAserotype5withdisulfidebondtype4,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminallipidation,C-terminalHistag(GLEHHHHHH) LipCSSFNGKGETSEKTIVRANGTRLEYTDIKSDGSGKAKEVLKDFTLEGTLAADGKTTLKVTCGTVVL SKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQKTKNLVFTKEDTITVQRYDSAGTNLE GKAVEITTLKELCNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEISEKTIVRACGTRLEYTDIKSDKT GKAKEVLKDFTLEGTLAADGKTTLKVTEGTVTLSKNISKSGEITVALDDTDSSGNKKSGTWDSGTSTL TISKNRTKTKQLVFTKECTITVQNYDSAGTNLEGKAVEITTLKELKNALKGLEHHHHHH SEQIDNO:138 Lip-S6D1-55D4_His_nt:CodingsequenceforheterodimerfusionproteinofOspAserotype6with disulfidebondtype1andOspAserotype5withdisulfidebondtype4,E.coliIpplipidationsignal, N-terminalCSSforadditionoflipids,LN1linkersequence,C-terminalHistag(GLEHHHHHH) ATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGCTGGCAGGTTGCT CAAGCTTCAACGGCAAAGGTGAAACGAGCGAAAAGACCATCGTGCGTGCGAACGGTACCCGCC TGGAATATACGGACATTAAATCGGACGGCAGCGGCAAAGCAAAGGAAGTCCTGAAAGATTTTAC GCTGGAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACGTGCGGCACCGTGGT TCTGTCAAAAAACATTCTGAAGTCGGGTGAAATCACCGCAGCTCTGGATGACAGCGATACCACG CGTGCTACGAAAAAGACCGGTAAATGGGATAGCAAGACCTCTACGCTGACCATTAGTGTCAACT CCCAGAAAACGAAGAATCTGGTGTTCACCAAAGAAGATACGATCACCGTTCAACGCTATGACAGT GCGGGCACCAACCTGGAAGGCAAAGCCGTTGAAATTACCACGCTGAAAGAACTGTGTAATGCTC TGAAAGGTACTAGTGACAAAAACAATGGCTCTGGTAGCAAAGAGAAAAACAAAGATGGCAAGTAC TCATTCAACGAAAAAGGCGAAATCAGTGAAAAAACCATTGTGCGTGCGTGTGGCACCCGTCTGG AATATACCGACATCAAGAGCGATAAAACGGGTAAAGCGAAGGAAGTTCTGAAAGATTTTACGCTG GAAGGTACCCTGGCAGCAGACGGTAAAACCACGCTGAAGGTGACCGAAGGTACCGTTACGCTG TCCAAAAACATTAGTAAGTCCGGCGAAATCACGGICGCCCTGGATGACACCGATAGCTCTGGCA ACAAAAAGAGCGGTACCTGGGACTCAGGCACCTCGACGCTGACCATTTCTAAAAATCGTACGAA AACCAAGCAGCTGGTCTTCACGAAAGAATGCACGATCACCGTGCAAAACTATGATAGCGCAGGT ACCAATCTGGAAGGCAAAGCTGTGGAAATTACCACGCTGAAAGAACTGAAGAATGCTCTGAAAG GTCTCGAGCACCACCACCACCACCAC SEQIDNO:140 Lip-S2D0-His:aminoacidsofpositions131-273ofBorreliaafzeliistrainK78,OspAserotype2,wild- typesequence,N-terminalCKQNforadditionoflipids,C-terminalHistag(GLEHHHHHH) LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELKNALKGLEHHHHHH SEQIDNO:141 Lip-S2D1-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype1 (aa182and269),N-terminalCKQNforadditionoflipids,C-terminalHistag(GLEHHHHHH) LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKCGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELCNALKGLEHHHHHH SEQIDNO:142 Lip-S2D2-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype2 (aa182and272),N-terminalCKQNforadditionoflipids,C-terminalHistag(GLEHHHHHH) LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKCGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELKNACKGLEHHHHHH SEQIDNO:143 Lip-S2D3-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype3 (aa244and259),N-terminalCKQNforadditionoflipids,C-terminalHistag(GLEHHHHHH) LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTICVQKYDSAGTNLEGTC VEIKTLDELKNALKGLEHHHHHH SEQIDNO:144 Lip-S2D4-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype4 (aa141and241),N-terminalCKQNforadditionoflipids,C-terminalHistag(GLEHHHHHH) LipCKQNELSAKTMTRECGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQCTITVQKYDSAGTNLEGTA VEIKTLDELKNALKGLEHHHHHH SEQIDNO:145 Lip-S2D5-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype5 (aa165and265),N-terminalCKQNforadditionoflipids,C-terminalHistag(GLEHHHHHH) LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNCTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTCDELKNALKGLEHHHHHH SEQIDNO:146 Lip-S2D6-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype6 (aa185and272),N-terminalCKQNforadditionoflipids,C-terminalHistag(GLEHHHHHH) LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTCTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELKNACKGLEHHHHHH SEQIDNO:147 Lip-S2D7-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype7 (aa199and223),N-terminalCKQNforadditionoflipids,C-terminalHistag(GLEHHHHHH) LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTCALNDTNTTQATKKTGAWDSKTSTCTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELKNALKGLEHHHHHH SEQIDNO:148 Lip-S2D8-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype8 (aa243and262),N-terminalCKQNforadditionoflipids,C-terminalHistag(GLEHHHHHH) LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTCTVQKYDSAGTNLEGT AVECKTLDELKNALKGLEHHHHHH SEQIDNO:149 Lip-S2D9-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype9 (aa184and204),N-terminalCKQNforadditionoflipids,C-terminalHistag(GLEHHHHHH) LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGCVTLSKEI AKSGEVTVALNDCNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELKNALKGLEHHHHHH SEQIDNO:150 Lip-S2D10-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype10 (aa201and214),N-terminalCKQNforadditionoflipids,C-terminalHistag(GLEHHHHHH) LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTVACNDTNTTQATKKTCAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELKNALKGLEHHHHHH SEQIDNO:151 Lip-S2D11-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype11 (aa246and259),N-terminalCKQNforadditionoflipids,C-terminalHistag(GLEHHHHHH) LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVCKYDSAGTNLEGTC VEIKTLDELKNALKGLEHHHHHH SEQIDNO:152 Lip-S2D12-His:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype12 (aa167and178),N-terminalCKQNforadditionoflipids,C-terminalHistag(GLEHHHHHH) LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTCEGKVANDKVTCEVKEGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELKNALKGLEHHHHHH SEQIDNO:153 Lip-S2D0:aminoacidsofpositions131-273ofBorreliaafzeliistrainK78,OspAserotype2,wild-type sequence,N-terminalCKQNforadditionoflipids LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELKNALK SEQIDNO:154 Lip-S2D1:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype1(aa 182and269),N-terminalCKQNforadditionoflipids LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKCGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELCNALK SEQIDNO:155 Lip-S2D2:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype2(aa 182and272),N-terminalCKQNforadditionoflipids LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKCGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELKNACK SEQIDNO:156 Lip-S2D3:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype3(aa 244and259),N-terminalCKQNforadditionoflipids LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTICVQKYDSAGTNLEGTC VEIKTLDELKNALK SEQIDNO:157 Lip-S2D4:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype4(aa 141and241),N-terminalCKQNforadditionoflipids LipCKQNELSAKTMTRECGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQCTITVQKYDSAGTNLEGTA VEIKTLDELKNALK SEQIDNO:158 Lip-S2D5:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype5(aa 165and265),N-terminalCKQNforadditionoflipids LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNCTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTCDELKNALK SEQIDNO:159 Lip-S2D6:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype6(aa 185and272),N-terminalCKQNforadditionoflipids LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTCTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELKNACK SEQIDNO:160 Lip-S2D7:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype7(aa 199and223),N-terminalCKQNforadditionoflipids LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTCALNDTNTTQATKKTGAWDSKTSTCTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELKNALK SEQIDNO:161 Lip-S2D8:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype8(aa 243and262),N-terminalCKQNforadditionoflipids LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTCTVQKYDSAGTNLEGT AVECKTLDELKNALK SEQIDNO:162 Lip-S2D9:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype9(aa 184and204),N-terminalCKQNforadditionoflipids LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGCVTLSKEI AKSGEVTVALNDCNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELKNALK SEQIDNO:163 Lip-S2D10:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype10(aa 201and214),N-terminalCKQNforadditionoflipids LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTVACNDINTTQATKKTCAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELKNALK SEQIDNO:164 Lip-S2D11:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype11(aa 246and259),N-terminalCKQNforadditionoflipids LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVCKYDSAGTNLEGTC VEIKTLDELKNALK SEQIDNO:165 Lip-S2D12:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype12(aa 167and178),N-terminalCKQNforadditionoflipids LipCKQNELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTCEGKVANDKVTCEVKEGTVTLSKEI AKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTA VEIKTLDELKNALK SEQIDNO:166 S2D0:aminoacidsofpositions131-273ofBorreliaafzeliistrainK78,OspAserotype2,wild-type sequence ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LKNALK SEQIDNO:167 S2D1:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype1(aa182 and269) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKCGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LCNALK SEQIDNO:168 S2D2:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype2(aa182 and272) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKCGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LKNACK SEQIDNO:169 S2D3:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype3(aa244 and259) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTICVQKYDSAGTNLEGTCVEIKTLDE LKNALK SEQIDNO:170 S2D4:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype4(aa141 and241) ELSAKTMTRECGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQCTITVQKYDSAGTNLEGTAVEIKTLDE LKNALK SEQIDNO:171 S2D5:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype5(aa165 and265) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNCTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTCDE LKNALK SEQIDNO:172 S2D6:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype6(aa185 and272) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTCTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LKNACK SEQIDNO:173 S2D7:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype7(aa199 and223) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT CALNDTNTTQATKKTGAWDSKTSTCTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LKNALK SEQIDNO:174 S2D8:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype8(aa243 and262) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTCTVQKYDSAGTNLEGTAVECKTLD ELKNALK SEQIDNO:175 S2D9:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype9(aa184 and204) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGCVTLSKEIAKSGEVT VALNDCNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LKNALK SEQIDNO:176 S2D10:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype10(aa201 and214) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VACNDTNTTQATKKTCAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDE LKNALK SEQIDNO:177 S2D11:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype11(aa246 and259) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVT VALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVCKYDSAGTNLEGTCVEIKTLDE LKNALK SEQIDNO:178 S2D12:aa131-273ofBorreliaafzeliistrainK78,OspAserotype2withdisulfidebondtype12(aa167 and178) ELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTCEGKVANDKVTCEVKEGTVTLSKEIAKSGEV TVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLD ELKNALK SEQIDNO:179 B.burgdorferis.s.(strainB31,serotype1),OspA_aa126-273withreplacedhLFA-likesequencefrom serotype1OspA FNEKGEVSEKIITRADGTRLEYTGIKSDGSGKAKEVLKNFTLEGKVANDKTTLVVKEGTVTLSKNISKS GEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSKKTKDLVFTKENTITVQQYDSNGTKLEGSAVEIT KLDEIKNALK SEQIDNO:180 B.garinii(strainPBr,serotype3),OspA_aa126-274 FNDKGKLSEKVVTRANGTRLEYTEIKNDGSGKAKEVLKGFALEGTLTDGGETKLTVTEGTVTLSKNIS KSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNSQKPKQLVFTKENTITVQNYNRAGNALEGSPA EIKDLAELKAALK SEQIDNO:181 B.bavariensis(strainPBi,serotype4),OspA_aa126-273 FNAKGELSEKTILRANGTRLEYTEIKSDGTGKAKEVLKDFALEGTLAADKTTLKVTEGTVVLSKHIPNS GEITVELNDSNSTQATKKTGKWDSNTSTLTISVNSKKTKNIVFTKEDTITVQKYDSAGTNLEGNAVEIK TLDELKNALK SEQIDNO:182 B.garinii(strainPHei,serotype5),OspA_aa126-273 FNEKGEISEKTIVRANGTRLEYTDIKSDKTGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVTLSKNISKS GEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRTKTKQLVFTKEDTITVQNYDSAGTNLEGKAVEITT LKELKNALK SEQIDNO:183 B.garinii(strainDK29,serotype6),OspA_aa126-274 FNGKGETSEKTIVRANGTRLEYTDIKSDGSGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVVLSKNILK SGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQKTKNLVFTKEDTITVQRYDSAGTNLEGKAVEI TTLKELKNALK SEQIDNO:184 LN1peptidelinkerconstructedfromtwoseparateloopregionsoftheN-terminalhalfofOspAfromB. burgdorferis.s.strainB31(aa65-74andaa42-53,aminoacidexchangeatposition53:D53S) GTSDKNNGSGSKEKNKDGKYS SEQIDNO:185 Lip-S1D4-S2D4_aa:HeterodimerfusionproteinofOspAserotypes1and2bothwithdisulfidebond type4,N-terminalCSSforadditionoflipids,LN1linkersequence,aa164-174ofOspAserotype1 replacedbynon-hLFA-1-likesequenceNFTLEGKVAND LipCSSFNEKGEVSEKIITRACGTRLEYTGIKSDGSGKAKEVLKNFTLEGKVANDKTTLVVKEGTVTLSK NISKSGEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSKKTKDLVFTKECTITVQQYDSNGTKLEG SAVEITKLDEIKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGELSAKTMTRECGTKLEYTEMKSDGT GKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTL TISVNSKKTTQLVFTKQCTITVQKYDSAGTNLEGTAVEIKTLDELKNALK SEQIDNO:186 Lip-S1D1-S2D1_aa:HeterodimerfusionproteinofOspAserotype1andOspAserotype2with disulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linkersequence,aa164-174of OspAserotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND,N-terminallipidation LipCSSFNEKGEVSEKIITRADGTRLEYTGIKSDGSGKAKEVLKNFTLEGKVANDKTTLVVKCGTVTLSK NISKSGEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSKKTKDLVFTKENTITVQQYDSNGTKLEG SAVEITKLDEICNALKGTSDKNNGSGSKEKNKDGKYSFNEKGELSAKTMTRENGTKLEYTEMKSDGT GKAKEVLKNFTLEGKVANDKVTLEVKCGTVTLSKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTL TISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDELCNALK SEQIDNO:187 Lip-S3D4-S4D4_aa:HeterodimerfusionproteinofOspAserotype3andOspAserotype4with disulfidebondtype4,N-terminalCSSforadditionoflipids,LN1linkersequence,N-terminal lipidation LipCSSFNEKGKLSEKVVTRACGTRLEYTEIKNDGSGKAKEVLKGFALEGTLTDGGETKLTVTEGTVTL SKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNSQKPKQLVFTKECTITVQNYNRAGNALE GSPAEIKDLAELKAALKGTSDKNNGSGSKEKNKDGKYSFNAKGELSEKTILRACGTRLEYTEIKSDGT GKAKEVLKDFALEGTLAADKTTLKVTEGTVVLSKHIPNSGEITVELNDSNSTQATKKTGKWDSNTSTL TISVNSKKTKNIVFTKECTITVQKYDSAGTNLEGNAVEIKTLDELKNALK SEQIDNO:188 Lip-S3D1-S4D1_aa:HeterodimerfusionproteinofOspAserotypes3and4bothwithdisulfidebond type1,N-terminalCSSforadditionoflipids,LN1linkersequence,N-terminallipidation LipCSSFNEKGKLSEKVVTRANGTRLEYTEIKNDGSGKAKEVLKGFALEGTLTDGGETKLTVTCGTVTL SKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNSQKPKQLVFTKENTITVQNYNRAGNALE GSPAEIKDLAELCAALKGTSDKNNGSGSKEKNKDGKYSFNAKGELSEKTILRANGTRLEYTEIKSDGT GKAKEVLKDFALEGTLAADKTTLKVTCGTVVLSKHIPNSGEITVELNDSNSTQATKKTGKWDSNTSTL TISVNSKKTKNIVFTKEDTITVQKYDSAGTNLEGNAVEIKTLDELCNALK SEQIDNO:189 Lip-S5D4-S6D4_aa:HeterodimerfusionproteinOspAserotypes5and6bothwithdisulfidebondtype 4,N-terminalCSSforadditionoflipids,LN1linkersequence,N-terminallipidation LipCSSFNEKGEISEKTIVRACGTRLEYTDIKSDKTGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVTLS KNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRTKTKQLVFTKECTITVQNYDSAGTNLEGK AVEITTLKELKNALKGTSDKNNGSGSKEKNKDGKYSFNGKGETSEKTIVRACGTRLEYTDIKSDGSGK AKEVLKDFTLEGTLAADGKTTLKVTEGTVVLSKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTIS VNSQKTKNLVFTKECTITVQRYDSAGTNLEGKAVEITTLKELKNALK SEQIDNO:190 Lip-S5D1-S6D1_aa:HeterodimerfusionproteinofOspAserotypes6bothwithdisulfidebondtype1, N-terminalCSSforadditionoflipids,LN1linkersequence,N-terminallipidation LipCSSFNEKGEISEKTIVRANGTRLEYTDIKSDKTGKAKEVLKDFTLEGTLAADGKTTLKVTCGTVTLS KNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRTKTKQLVFTKEDTITVQNYDSAGTNLEGK AVEITTLKELCNALKGTSDKNNGSGSKEKNKDGKYSFNGKGETSEKTIVRANGTRLEYTDIKSDGSGK AKEVLKDFTLEGTLAADGKTTLKVTCGTVVLSKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTIS VNSQKTKNLVFTKEDTITVQRYDSAGTNLEGKAVEITTLKELCNALK SEQIDNO:191 Lip-S2D4-S1D4_aa:HeterodimerfusionproteinofOspAserotypes2and1bothwithdisulfidebond type4,N-terminalCSSforadditionoflipids,LN1linkersequence,aa164-174ofOspAserotype1 replacedbynon-hLFA-1-likesequenceNFTLEGKVAND,N-terminallipidation LipCSSFNEKGELSAKTMTRECGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTL SKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQCTITVQKYDSAGTNLE GTAVEIKTLDELKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEVSEKIITRACGTRLEYTGIKSDGS GKAKEVLKNFTLEGKVANDKTTLVVKEGTVTLSKNISKSGEVSVELNDTDSSAATKKTAAWNSGTSTL TITVNSKKTKDLVFTKECTITVQQYDSNGTKLEGSAVEITKLDEIKNALK SEQIDNO:192 Lip-S2D1-S1D1_aa:HeterodimerfusionproteinofOspAserotypes2and1bothwithdisulfidebond type1,N-terminalCSSforadditionoflipids,LN1linkersequence,aa164-174ofOspAserotype1 replacedbynon-hLFA-1-likesequenceNFTLEGKVAND,N-terminallipidation LipCSSFNEKGELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKCGTVTL SKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLE GTAVEIKTLDELCNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEVSEKIITRADGTRLEYTGIKSDGS GKAKEVLKNFTLEGKVANDKTTLVVKCGTVTLSKNISKSGEVSVELNDTDSSAATKKTAAWNSGTSTL TITVNSKKTKDLVFTKENTITVQQYDSNGTKLEGSAVEITKLDEICNALK SEQIDNO:193 Lip-S4D4-S3D4_aa:HeterodimerfusionproteinofOspAserotypes4and3bothwithdisulfidebond type4,N-terminalCSSforadditionoflipids,LN1linkersequence,N-terminallipidation LipCSSFNAKGELSEKTILRACGTRLEYTEIKSDGTGKAKEVLKDFALEGTLAADKTTLKVTEGTVVLSK HIPNSGEITVELNDSNSTQATKKTGKWDSNTSTLTISVNSKKTKNIVFTKECTITVQKYDSAGTNLEGN AVEIKTLDELKNALKGTSDKNNGSGSKEKNKDGKYSFNDKGKLSEKVVTRACGTRLEYTEIKNDGSG KAKEVLKGFALEGTLTDGGETKLTVTEGTVTLSKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLT ISKNSQKPKQLVFTKECTITVQNYNRAGNALEGSPAEIKDLAELKAALK SEQIDNO:194 Lip-S4D1-S3D1_aa:HeterodimerfusionproteinofOspAserotypes4and3bothwithdisulfidebond type1,N-terminalCSSforadditionoflipids,LN1linkersequence,N-terminallipidation LipCSSFNAKGELSEKTILRANGTRLEYTEIKSDGTGKAKEVLKDFALEGTLAADKTTLKVTCGTVVLSK HIPNSGEITVELNDSNSTQATKKTGKWDSNTSTLTISVNSKKTKNIVFTKEDTITVQKYDSAGTNLEGN AVEIKTLDELCNALKGTSDKNNGSGSKEKNKDGKYSFNDKGKLSEKVVTRANGTRLEYTEIKNDGSG KAKEVLKGFALEGTLTDGGETKLTVTCGTVTLSKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLT ISKNSQKPKQLVFTKENTITVQNYNRAGNALEGSPAEIKDLAELCAALK SEQIDNO:195 Lip-S6D4-S5D4_aa:HeterodimerfusionproteinofOspAserotypes6and5bothwithdisulfidebond type4,N-terminalCSSforadditionoflipids,LN1linkersequence,N-terminallipidation LipCSSFNGKGETSEKTIVRACGTRLEYTDIKSDGSGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVVL SKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQKTKNLVFTKECTITVQRYDSAGTNLE GKAVEITTLKELKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEISEKTIVRACGTRLEYTDIKSDKTG KAKEVLKDFTLEGTLAADGKTTLKVTEGTVTLSKNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTI SKNRTKTKQLVFTKECTITVQNYDSAGTNLEGKAVEITTLKELKNALK SEQIDNO:196 Lip-S6D1-S5D1_aa:HeterodimerfusionproteinofOspAserotypes6and5bothwithdisulfidebond type1,N-terminalCSSforadditionoflipids,LN1linkersequence,N-terminallipidation LipCSSFNGKGETSEKTIVRANGTRLEYTDIKSDGSGKAKEVLKDFTLEGTLAADGKTTLKVTCGTVVL SKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQKTKNLVFTKEDTITVQRYDSAGTNLE GKAVEITTLKELCNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEISEKTIVRANGTRLEYTDIKSDKT GKAKEVLKDFTLEGTLAADGKTTLKVTCGTVTLSKNISKSGEITVALDDTDSSGNKKSGTWDSGTSTL TISKNRTKTKQLVFTKEDTITVQNYDSAGTNLEGKAVEITTLKELCNALK SEQIDNO:197 Lip-S1D4-S2D1_aa:HeterodimerfusionproteinofOspAserotype1withdisulfidebondtype4and OspAserotypewithdisulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linker Isequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND, N-terminallipidation LipCSSFNEKGEVSEKIITRACGTRLEYTGIKSDGSGKAKEVLKNFTLEGKVANDKTTLVVKEGTVTLSK NISKSGEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSKKTKDLVFTKECTITVQQYDSNGTKLEG SAVEITKLDEIKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGELSAKTMTRENGTKLEYTEMKSDGT GKAKEVLKNFTLEGKVANDKVTLEVKCGTVTLSKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTL TISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDELCNALK SEQIDNO:198 Lip-S1D1-S2D4_aa:HeterodimerfusionproteinofOspAserotype1withdisulfidebondtype1and OspAserotype2withdisulfidebondtype4,N-terminalCSSforadditionoflipids,LN1linker sequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND, N-terminallipidation LipCSSFNEKGEVSEKIITRADGTRLEYTGIKSDGSGKAKEVLKNFTLEGKVANDKTTLVVKCGTVTLSK NISKSGEVSVELNDTDSSAATKKTAAWNSGTSTLTITVNSKKTKDLVFTKENTITVQQYDSNGTKLEG SAVEITKLDEICNALKGTSDKNNGSGSKEKNKDGKYSFNEKGELSAKTMTRECGTKLEYTEMKSDGT GKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTL TISVNSKKTTQLVFTKQCTITVQKYDSAGTNLEGTAVEIKTLDELKNALK SEQIDNO:199 Lip-S3D4-S4D1_aa:HeterodimerfusionproteinofOspAserotype3withdisulfidebondtype4and OspAserotype4withdisulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminallipidation LipCSSFNEKGKLSEKVVTRACGTRLEYTEIKNDGSGKAKEVLKGFALEGTLTDGGETKLTVTEGTVTL SKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNSQKPKQLVFTKECTITVQNYNRAGNALE GSPAEIKDLAELKAALKGTSDKNNGSGSKEKNKDGKYSFNAKGELSEKTILRANGTRLEYTEIKSDGT GKAKEVLKDFALEGTLAADKTTLKVTCGTVVLSKHIPNSGEITVELNDSNSTQATKKTGKWDSNTSTL TISVNSKKTKNIVFTKEDTITVQKYDSAGTNLEGNAVEIKTLDELCNALK SEQIDNO:200 Lip-S3D1-S4D4_aa:HeterodimerfusionproteinofOspAserotype3withdisulfidebondtype1and OspAserotype4withdisulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminallipidation LipCSSFNEKGKLSEKVVTRANGTRLEYTEIKNDGSGKAKEVLKGFALEGTLTDGGETKLTVTCGTVTL SKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLTISKNSQKPKQLVFTKENTITVQNYNRAGNALE GSPAEIKDLAELCAALKGTSDKNNGSGSKEKNKDGKYSFNAKGELSEKTILRACGTRLEYTEIKSDGT GKAKEVLKDFALEGTLAADKTTLKVTEGTVVLSKHIPNSGEITVELNDSNSTQATKKTGKWDSNTSTL TISVNSKKTKNIVFTKECTITVQKYDSAGTNLEGNAVEIKTLDELKNALK SEQIDNO:201 Lip-S5D4-S6D1_aa:HeterodimerfusionproteinofOspAserotype5withdisulfidebondtype4and OspAserotype6withdisulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminallipidation LipCSSFNEKGEISEKTIVRACGTRLEYTDIKSDKTGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVTLS KNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRTKTKQLVFTKECTITVQNYDSAGTNLEGK AVEITTLKELKNALKGTSDKNNGSGSKEKNKDGKYSFNGKGETSEKTIVRANGTRLEYTDIKSDGSGK AKEVLKDFTLEGTLAADGKTTLKVTCGTVVLSKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTIS VNSQKTKNLVFTKEDTITVQRYDSAGTNLEGKAVEITTLKELCNALK SEQIDNO:202 Lip-S5D1-S6D4_aa:HeterodimerfusionproteinofOspAserotype5withdisulfidebondtype1and OspAserotype6withdisulfidebondtype4,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminallipidation LipCSSFNEKGEISEKTIVRANGTRLEYTDIKSDKTGKAKEVLKDFTLEGTLAADGKTTLKVTCGTVTLS KNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTISKNRTKTKQLVFTKEDTITVQNYDSAGTNLEGK AVEITTLKELCNALKGTSDKNNGSGSKEKNKDGKYSFNGKGETSEKTIVRACGTRLEYTDIKSDGSGK AKEVLKDFTLEGTLAADGKTTLKVTEGTVVLSKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTIS VNSQKTKNLVFTKECTITVQRYDSAGTNLEGKAVEITTLKELKNALK SEQIDNO:203 Lip-S2D4-S1D1_aa:HeterodimerfusionproteinofOspAserotype2withdisulfidebondtype4and OspAserotype1withdisulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linker sequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND, N-terminallipidation LipCSSFNEKGELSAKTMTRECGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTL SKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQCTITVQKYDSAGTNLE GTAVEIKTLDELKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEVSEKIITRADGTRLEYTGIKSDGS GKAKEVLKNFTLEGKVANDKTTLVVKCGTVTLSKNISKSGEVSVELNDTDSSAATKKTAAWNSGTSTL TITVNSKKTKIDLVFTKENTITVQQYDSNGTKLEGSAVEITKLDEICNALK SEQIDNO:204 Lip-S2D1-S1D4_aa:HeterodimerfusionproteinofOspAserotype2withdisulfidebondtype1and OspAserotype1withdisulfidebondtype4,N-terminalCSSforadditionoflipids,LN1linker sequence,aa164-174ofOspAserotype1replacedbynon-hLFA-1-likesequenceNFTLEGKVAND, N-terminallipidation LipCSSFNEKGELSAKTMTRENGTKLEYTEMKSDGTGKAKEVLKNFTLEGKVANDKVTLEVKCGTVTL SKEIAKSGEVTVALNDTNTTQATKKTGAWDSKTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLE GTAVEIKTLDELCNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEVSEKIITRACGTRLEYTGIKSDGS GKAKEVLKNFTLEGKVANDKTTLVVKEGTVTLSKNISKSGEVSVELNDTDSSAATKKTAAWNSGTSTL TITVNSKKTKDLVFTKECTITVQQYDSNGTKLEGSAVEITKLDEIKNALK SEQIDNO:205 Lip-S4D4-S3D1_aa:HeterodimerfusionproteinofOspAserotype4withdisulfidebondtype4and OspAserotype3withdisulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminallipidation LipCSSFNAKGELSEKTILRACGTRLEYTEIKSDGTGKAKEVLKDFALEGTLAADKTTLKVTEGTVVLSK HIPNSGEITVELNDSNSTQATKKTGKWDSNTSTLTISVNSKKTKNIVFTKECTITVQKYDSAGTNLEGN AVEIKTLDELKNALKGTSDKNNGSGSKEKNKDGKYSFNDKGKLSEKVVTRANGTRLEYTEIKNDGSG KAKEVLKGFALEGTLTDGGETKLTVTCGTVTLSKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLT ISKNSQKPKQLVFTKENTITVQNYNRAGNALEGSPAEIKDLAELCAALK SEQIDNO:206 Lip-S4D1-S3D4_aa:CodingsequenceforintermediateandfinalheterodimerfusionproteinsofOspA serotype4withdisulfidebondtype1andOspAserotype3withdisulfidebondtype4,N-terminalCSS foradditionoflipids,LN1linkersequence,N-terminallipidation LipCSSFNAKGELSEKTILRANGTRLEYTEIKSDGTGKAKEVLKDFALEGTLAADKTTLKVTCGTVVLSK HIPNSGEITVELNDSNSTQATKKTGKWDSNTSTLTISVNSKKTKNIVFTKEDTITVQKYDSAGTNLEGN AVEIKTLDELCNALKGTSDKNNGSGSKEKNKDGKYSFNDKGKLSEKVVTRACGTRLEYTEIKNDGSG KAKEVLKGFALEGTLTDGGETKLTVTEGTVTLSKNISKSGEITVALNDTETTPADKKTGEWKSDTSTLT ISKNSQKPKQLVFTKECTITVQNYNRAGNALEGSPAEIKDLAELKAALK SEQIDNO:207 Lip-S6D4-S5D1_aa:HeterodimerfusionproteinofOspAserotype6withdisulfidebondtype4and OspAserotype5withdisulfidebondtype1,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminallipidation LipCSSFNGKGETSEKTIVRACGTRLEYTDIKSDGSGKAKEVLKDFTLEGTLAADGKTTLKVTEGTVVL SKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQKTKNLVFTKECTITVQRYDSAGTNLE GKAVEITTLKELKNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEISEKTIVRANGTRLEYTDIKSDKTG KAKEVLKDFTLEGTLAADGKTTLKVTCGTVTLSKNISKSGEITVALDDTDSSGNKKSGTWDSGTSTLTI SKNRTKTKQLVFTKEDTITVQNYDSAGTNLEGKAVEITTLKELCNALK SEQIDNO:208 Lip-S6D1-S5D4_aa:HeterodimerfusionproteinofOspAserotype6withdisulfidebondtype1and OspAserotype5withdisulfidebondtype4,N-terminalCSSforadditionoflipids,LN1linker sequence,N-terminallipidation LipCSSFNGKGETSEKTIVRANGTRLEYTDIKSDGSGKAKEVLKDFTLEGTLAADGKTTLKVTCGTVVL SKNILKSGEITAALDDSDTTRATKKTGKWDSKTSTLTISVNSQKTKNLVFTKEDTITVQRYDSAGTNLE GKAVEITTLKELCNALKGTSDKNNGSGSKEKNKDGKYSFNEKGEISEKTIVRACGTRLEYTDIKSDKT GKAKEVLKDFTLEGTLAADGKTTLKVTEGTVTLSKNISKSGEITVALDDTDSSGNKKSGTWDSGTSTL TISKNRTKTKQLVFTKECTITVQNYDSAGTNLEGKAVEITTLKELKNALK SEQIDNO:209 B.afzelii(strainK78;OspAserotype2)aa17-273,lipidationsignalsequenceremoved(aa1-16: MKKYLLGIGLILALIA),C-terminalHistag(GLEHHHHHH) CKQNVSSLDEKNSASVDLPGEMKVLVSKEKDKDGKYSLKATVDKIELKGTSDKDNGSGVLEGTKDDK SKAKLTIADDLSKTTFELFKEDGKTLVSRKVSSKDKTSTDEMFNEKGELSAKTMTRENGTKLEYTEMK SDGTGKAKEVLKNFTLEGKVANDKVTLEVKEGTVTLSKEIAKSGEVTVALNDTNTTQATKKTGAWDS KTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDELKNALKGLEHHHHHH SEQIDNO:210 B.burgdorferi(OspAserotype1,strainZS7)aa17-273,lipidationsignalsequenceremoved(aa1-16: MKKYLLGIGLILALIA),C-terminalHistag(LEHHHHHH) CSSFKQNVSSLDEKNSVSVDLPGEMKVLVSKEKNKDGKYDLIATVDKLELKGTSDKNNGSGVLEGVK ADKSKVKLTISDDLGQTTLEVFKEDGKTLVSKKVTSKDKSSTEEKFNEKGEVSEKIITRADGTRLEYTGI KSDGSGKAKEVLKGYVLEGTLTAEKTTLVVKEGTVTLSKNISKSGEVSVELNDTDSSAATKKTAAWNS GTSTLTITVNSKKTKDLVFTKENTITVQQYDSNGTKLEGSAVEITKLDEIKNALKLEHHHHHH SEQIDNO:211 Cysteine-containingpeptidefromOspA CKQN SEQIDNO:212 ChimericOspASerotype1/Serotype2,N-terminallipidation LipCAQKGAESIGSVSVDLPGEMKVLVSKEKDKNGKYDLIATVDKLELKGTSDKNNGSGVLEGVKTNK SKVKLTISDDLGQTTLEVFKEDGKTLVSKKVTSKDKSSTEEKFNEKGEVSEKIITMADGTRLEYTGIKS DGTGKAKYVLKNFTLEGKVANDKTTLEVKEGTVTLSMNISKSGEVSVELNDTDSSAATKKTAAWNSK TSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGTNLEGTAVEIKTLDELKNALKLE SEQIDNO:213 aminoacidsofpositions126-274ofB.garinfistrainT25,OspAserotype7 FNDKGKLSEKVVTRANGTRLEYTEIQNDGSGKAKEVLKSLTLEGTLTADGETKLTVEAGTVTLSKNISE SGEITVELKDTETTPADKKSGTWDSKTSTLTISKNSQKTKQLVFTKENTITVQKYNTAGTKLEGSPAEI KDLEALKAALK SEQIDNO:214 ForwardoligonucleotideprimerfortheRecAgeneofBorrelia CATGCTCTTGATCCTGTTTA SEQIDNO:215 Histidinetag GLEHHHHHH SEQIDNO:216 ReverseoligonucleotideprimerfortheRecAgeneofBorrelia CCCATTTCTCCATCTATCTC

[0447] The entire contents of all of the references (including literature references, issued patents, published patent applications, and co-pending patent applications) cited throughout this application are hereby expressly incorporated by reference.