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. 1991 Jun;59(6):2070–2074. doi: 10.1128/iai.59.6.2070-2074.1991

Further characterization of a potent immunogen and the chromosomal gene encoding it in the Lyme disease agent, Borrelia burgdorferi.

G C Perng 1, R B LeFebvre 1, R C Johnson 1
PMCID: PMC257967  PMID: 2037367

Abstract

Further characterization of a previously reported 83-kDa antigen of Borrelia burgdorferi and the gene encoding it is reported. The DNA sequence of the gene and the amino acid sequence of the protein were determined. On the basis of the amino acid content, the actual size of the antigen was determined to be 79.8 kDa, rather than 83 kDa as previously reported. The expression of the antigen in several representative North American and European B. burgdorferi isolates was demonstrated. The conservation of the gene within the species was demonstrated by DNA hybridization of a labeled gene probe to several North American and European B. burgdorferi isolates. Of other Borrelia spp. assayed (B. hermsii, B. coriaceae, B. duttonii, and B. anserina), only B. anserina, a poultry pathogen, hybridized to the gene probe and expressed the 79.8-kDa antigen.

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Selected References

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  1. Anderson J. F., Magnarelli L. A., LeFebvre R. B., Andreadis T. G., McAninch J. B., Perng G. C., Johnson R. C. Antigenically variable Borrelia burgdorferi isolated from cottontail rabbits and Ixodes dentatus in rural and urban areas. J Clin Microbiol. 1989 Jan;27(1):13–20. doi: 10.1128/jcm.27.1.13-20.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson J. F., Magnarelli L. A., McAninch J. B. New Borrelia burgdorferi antigenic variant isolated from Ixodes dammini from upstate New York. J Clin Microbiol. 1988 Oct;26(10):2209–2212. doi: 10.1128/jcm.26.10.2209-2212.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barbour A. G., Garon C. F. Linear plasmids of the bacterium Borrelia burgdorferi have covalently closed ends. Science. 1987 Jul 24;237(4813):409–411. doi: 10.1126/science.3603026. [DOI] [PubMed] [Google Scholar]
  4. Barbour A. G., Heiland R. A., Howe T. R. Heterogeneity of major proteins in Lyme disease borreliae: a molecular analysis of North American and European isolates. J Infect Dis. 1985 Sep;152(3):478–484. doi: 10.1093/infdis/152.3.478. [DOI] [PubMed] [Google Scholar]
  5. Barbour A. G. Isolation and cultivation of Lyme disease spirochetes. Yale J Biol Med. 1984 Jul-Aug;57(4):521–525. [PMC free article] [PubMed] [Google Scholar]
  6. Barbour A. G. Plasmid analysis of Borrelia burgdorferi, the Lyme disease agent. J Clin Microbiol. 1988 Mar;26(3):475–478. doi: 10.1128/jcm.26.3.475-478.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Barbour A. G., Tessier S. L., Hayes S. F. Variation in a major surface protein of Lyme disease spirochetes. Infect Immun. 1984 Jul;45(1):94–100. doi: 10.1128/iai.45.1.94-100.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bissett M. L., Hill W. Characterization of Borrelia burgdorferi strains isolated from Ixodes pacificus ticks in California. J Clin Microbiol. 1987 Dec;25(12):2296–2301. doi: 10.1128/jcm.25.12.2296-2301.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bundoc V. G., Barbour A. G. Clonal polymorphisms of outer membrane protein OspB of Borrelia burgdorferi. Infect Immun. 1989 Sep;57(9):2733–2741. doi: 10.1128/iai.57.9.2733-2741.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Burgdorfer W., Barbour A. G., Hayes S. F., Benach J. L., Grunwaldt E., Davis J. P. Lyme disease-a tick-borne spirochetosis? Science. 1982 Jun 18;216(4552):1317–1319. doi: 10.1126/science.7043737. [DOI] [PubMed] [Google Scholar]
  11. Coleman J. L., Benach J. L. Isolation of antigenic components from the Lyme disease spirochete: their role in early diagnosis. J Infect Dis. 1987 Apr;155(4):756–765. doi: 10.1093/infdis/155.4.756. [DOI] [PubMed] [Google Scholar]
  12. Craft J. E., Grodzicki R. L., Steere A. C. Antibody response in Lyme disease: evaluation of diagnostic tests. J Infect Dis. 1984 May;149(5):789–795. doi: 10.1093/infdis/149.5.789. [DOI] [PubMed] [Google Scholar]
  13. Dattwyler R. J., Volkman D. J., Luft B. J. Immunologic aspects of Lyme borreliosis. Rev Infect Dis. 1989 Sep-Oct;11 (Suppl 6):S1494–S1498. doi: 10.1093/clinids/11.supplement_6.s1494. [DOI] [PubMed] [Google Scholar]
  14. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  15. Grodzicki R. L., Steere A. C. Comparison of immunoblotting and indirect enzyme-linked immunosorbent assay using different antigen preparations for diagnosing early Lyme disease. J Infect Dis. 1988 Apr;157(4):790–797. doi: 10.1093/infdis/157.4.790. [DOI] [PubMed] [Google Scholar]
  16. Hansen K., Bangsborg J. M., Fjordvang H., Pedersen N. S., Hindersson P. Immunochemical characterization of and isolation of the gene for a Borrelia burgdorferi immunodominant 60-kilodalton antigen common to a wide range of bacteria. Infect Immun. 1988 Aug;56(8):2047–2053. doi: 10.1128/iai.56.8.2047-2053.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hansen K., Hindersson P., Pedersen N. S. Measurement of antibodies to the Borrelia burgdorferi flagellum improves serodiagnosis in Lyme disease. J Clin Microbiol. 1988 Feb;26(2):338–346. doi: 10.1128/jcm.26.2.338-346.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hinnebusch J., Bergström S., Barbour A. G. Cloning and sequence analysis of linear plasmid telomeres of the bacterium Borrelia burgdorferi. Mol Microbiol. 1990 May;4(5):811–820. doi: 10.1111/j.1365-2958.1990.tb00651.x. [DOI] [PubMed] [Google Scholar]
  19. Howe T. R., LaQuier F. W., Barbour A. G. Organization of genes encoding two outer membrane proteins of the Lyme disease agent Borrelia burgdorferi within a single transcriptional unit. Infect Immun. 1986 Oct;54(1):207–212. doi: 10.1128/iai.54.1.207-212.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Howe T. R., Mayer L. W., Barbour A. G. A single recombinant plasmid expressing two major outer surface proteins of the Lyme disease spirochete. Science. 1985 Feb 8;227(4687):645–646. doi: 10.1126/science.3969554. [DOI] [PubMed] [Google Scholar]
  21. Jiang W., Luft B. J., Munoz P., Dattwyler R. J., Gorevic P. D. Cross-antigenicity between the major surface proteins (ospA and ospB) and other proteins of Borrelia burgdorferi. J Immunol. 1990 Jan 1;144(1):284–289. [PubMed] [Google Scholar]
  22. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  23. Lane R. S., Pascocello J. A. Antigenic characteristics of Borrelia burgdorferi isolates from ixodid ticks in California. J Clin Microbiol. 1989 Oct;27(10):2344–2349. doi: 10.1128/jcm.27.10.2344-2349.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. LeFebvre R. B., Lane R. S., Perng G. C., Brown J. A., Johnson R. C. DNA and protein analyses of tick-derived isolates of Borrelia burgdorferi from California. J Clin Microbiol. 1990 Apr;28(4):700–707. doi: 10.1128/jcm.28.4.700-707.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. LeFebvre R. B., Perng G. C., Johnson R. C. Characterization of Borrelia burgdorferi isolates by restriction endonuclease analysis and DNA hybridization. J Clin Microbiol. 1989 Apr;27(4):636–639. doi: 10.1128/jcm.27.4.636-639.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lefebvre R. B., Perng G. C., Johnson R. C. The 83-kilodalton antigen of Borrelia burgdorferi which stimulates immunoglobulin M (IgM) and IgG responses in infected hosts is expressed by a chromosomal gene. J Clin Microbiol. 1990 Jul;28(7):1673–1675. doi: 10.1128/jcm.28.7.1673-1675.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Muhlemann M. F., Wright D. J., Black C. Serology of Lyme disease. Lancet. 1986 Mar 8;1(8480):553–554. doi: 10.1016/s0140-6736(86)90903-7. [DOI] [PubMed] [Google Scholar]
  28. Nadal D., Taverna C., Hitzig W. H. Immunoblot analysis of antibody binding to polypeptides of Borrelia burgdorferi in children with different clinical manifestations of Lyme disease. Pediatr Res. 1989 Oct;26(4):377–382. doi: 10.1203/00006450-198910000-00019. [DOI] [PubMed] [Google Scholar]
  29. Pribnow D. Nucleotide sequence of an RNA polymerase binding site at an early T7 promoter. Proc Natl Acad Sci U S A. 1975 Mar;72(3):784–788. doi: 10.1073/pnas.72.3.784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  31. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  32. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schmid G. P., Steigerwalt A. G., Johnson S. E., Barbour A. G., Steere A. C., Robinson I. M., Brenner D. J. DNA characterization of the spirochete that causes Lyme disease. J Clin Microbiol. 1984 Aug;20(2):155–158. doi: 10.1128/jcm.20.2.155-158.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schwan T. G., Burgdorfer W. Antigenic changes of Borrelia burgdorferi as a result of in vitro cultivation. J Infect Dis. 1987 Nov;156(5):852–853. doi: 10.1093/infdis/156.5.852-a. [DOI] [PubMed] [Google Scholar]
  35. Schwan T. G., Burgdorfer W., Garon C. F. Changes in infectivity and plasmid profile of the Lyme disease spirochete, Borrelia burgdorferi, as a result of in vitro cultivation. Infect Immun. 1988 Aug;56(8):1831–1836. doi: 10.1128/iai.56.8.1831-1836.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  38. Steere A. C., Malawista S. E., Bartenhagen N. H., Spieler P. N., Newman J. H., Rahn D. W., Hutchinson G. J., Green J., Snydman D. R., Taylor E. The clinical spectrum and treatment of Lyme disease. Yale J Biol Med. 1984 Jul-Aug;57(4):453–461. [PMC free article] [PubMed] [Google Scholar]
  39. Steere A. C., Malawista S. E., Snydman D. R., Shope R. E., Andiman W. A., Ross M. R., Steele F. M. Lyme arthritis: an epidemic of oligoarticular arthritis in children and adults in three connecticut communities. Arthritis Rheum. 1977 Jan-Feb;20(1):7–17. doi: 10.1002/art.1780200102. [DOI] [PubMed] [Google Scholar]
  40. Steitz J. A. Polypeptide chain initiation: nucleotide sequences of the three ribosomal binding sites in bacteriophage R17 RNA. Nature. 1969 Dec 6;224(5223):957–964. doi: 10.1038/224957a0. [DOI] [PubMed] [Google Scholar]
  41. Thomas D. D., Comstock L. E. Interaction of Lyme disease spirochetes with cultured eucaryotic cells. Infect Immun. 1989 Apr;57(4):1324–1326. doi: 10.1128/iai.57.4.1324-1326.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Walker R. L., Greene R. T., Nicholson W. L., Levine J. F. Shared flagellar epitopes of Borrelia burgdorferi and Borrelia anserina. Vet Microbiol. 1989 Apr;19(4):361–371. doi: 10.1016/0378-1135(89)90101-6. [DOI] [PubMed] [Google Scholar]
  43. Wallich R., Moter S. E., Simon M. M., Ebnet K., Heiberger A., Kramer M. D. The Borrelia burgdorferi flagellum-associated 41-kilodalton antigen (flagellin): molecular cloning, expression, and amplification of the gene. Infect Immun. 1990 Jun;58(6):1711–1719. doi: 10.1128/iai.58.6.1711-1719.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wilske B., Preac-Mursic V., Schierz G., Kühbeck R., Barbour A. G., Kramer M. Antigenic variability of Borrelia burgdorferi. Ann N Y Acad Sci. 1988;539:126–143. doi: 10.1111/j.1749-6632.1988.tb31846.x. [DOI] [PubMed] [Google Scholar]
  45. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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