1887

Abstract

This study describes a PCR-based approach for the production of a rationally attenuated mutant of . Degenerate primers were used to amplify a fragment encoding 91.45% of the gene of MP6 which was cloned into pUC18. The remainder of the gene was isolated by inverse PCR. The gene was sequenced and a restriction map was generated. The gene had 75.9% identity with the gene of . The cloned gene was inactivated and reintroduced into strain GB using the suicide vector pGP704. A stable -defective mutant, GBΔ, was isolated and its virulence was examined . The mutant was attenuated in guinea-pigs and capable of inducing a protective immune response against challenge with the virulent strain GB. Unusually for an -defective mutant, the mutant was virulent in mice, with a median dose which induced morbidity or death similar to that of the wild-type, although time to death was significantly prolonged.

Keyword(s): aroA and Yersinia pestis
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1996-07-01
2021-05-19
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References

  1. Alexander J. E., Andrew P. W., Jones D., Roberts I. S. 1993; Characterization of an aromatic amino acid-dependent Listeria monocytogenes mutant : attenuation, persistence and ability to produce protective immunity in mice. Infect Immun 61:2245–2248
    [Google Scholar]
  2. Bowe F., O'Gaora P., Masked D., Cafferkey M., Dougan G. 1989; Virulence, persistence and immunogenicity of Yersinia enterocolitica 0:8 aro A mutants. Infect Immun 57:3234–3236
    [Google Scholar]
  3. Burrows T.W. 1960; Biochemical properties of virulent and avirulent strains of bacteria : Salmonella typhosa and Pasteurella pestis.. Ann NY Acad Sci 88:1125–1135
    [Google Scholar]
  4. Burrows T.W. 1963; Virulence of Pasteurella pestis and immunity to plague. In Ergebnisse der Mikrobiologie Immunitatsforschung und Experimentellen Therapie pp. 59–113 Berlin: Springer-Verlag;
    [Google Scholar]
  5. Burrows T.W., Gillet W. A. 1971; Host specificity of Brazilian strains of Pasteurella pestis. Nature 229:51–52
    [Google Scholar]
  6. Clements J. D., Bao J. X., Cardenas L. 1992; Use of attenuated bacteria as live oral vaccine vectors. In Recombinant DNA Vaccines: Rationale and Strategy pp. 293–322 Edited by Isaacson R. E. New York: Marcel Dekker;
    [Google Scholar]
  7. Cowling P., Moss P. 1994; Infectivity of pneumonic plague.. Br Med J 309:1369
    [Google Scholar]
  8. Davis K. J., Fritz D. L., Pitt M. L., Welkos S. L., Worsham P. L., Friedlander A. M. 1996; Pathology of experimental pneumonic plague produced by fraction 1-positive and fraction 1-negative Yersinia pestis in African green monkeys (Cercopitheus aethiops). Arch Pathol Lab Med 120:156–163
    [Google Scholar]
  9. Demidova G. V., Goncharov E. K., Tynyanova V. I. 1984; Comparison of the recognition site specificities of adenyl and cytosyl DNA methylases of Yersinia pestis EV76 dam and dem with E. coli methylases.. Biokhimiya 49:1594–1597
    [Google Scholar]
  10. Dower W. J., Miller J. F., Ragsdale C. W. 1988; High efficiency transformation of E. coti by high voltage electroporation. Nucleic Acids Res 16:6127–6145
    [Google Scholar]
  11. Drozdov I., Anisimov A. P., Samoilova L. V., Yezhov I. N., Yeremin S. A., Karlyshev A. V., Krasilnikova V. M., Kravchenko V. I. 1995; Virulent non-capsulate Yersinia pestis variants constructed by insertion mutagenesis.. J Med Microbiol 42:264–268
    [Google Scholar]
  12. Duncan K., Coggins J. R. 1986; The ser C-aro A operon of Escherichia coli.. Biochem J 234:49–57
    [Google Scholar]
  13. Dybvig K., Hollinshead S. K., Heath D. G., Clewell D. B., Sun F., Woodard A. 1992; Degenerate oligonucleotide primers for enzymatic amplification of rec A sequences from Gram positive bacteria and mycobacteria. J Bacteriol 174:2729–2732
    [Google Scholar]
  14. Grinter N.J. 1983; A broad-host-range cloning vector trans- posable to various replicons. Gene 21:133–143
    [Google Scholar]
  15. Havard H. L., Hunter S. E. C., Titball R. W. 1992; Comparison of the nucleotide sequence and development of a PCR test for the epsilon toxin gene of Clostridium perfringens.. FEMS Lett 97:77–82
    [Google Scholar]
  16. Heesemann J., Hantke K., Vocke T., Saken E., Rakin A., Stojiljkovic I., Berner R. 1993; Virulence of Yersinia enterocolitica is closely associated with siderophore production, expression of an iron-repressible outer membrane polypeptide of 65,000 Da and pesticin sensitivity. Mol Microbiol 8:397–408
    [Google Scholar]
  17. Hoiseth S.K., Stocker B. A. D. 1981; Aromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines. Nature 291:238–239
    [Google Scholar]
  18. Homchampa P., Strugnell R. A., Adler B. 1992; Molecular analysis of the aro A gene of Pasteurella multocida and vaccine potential of a constructed aro A mutant. Mol Microbiol 6:3585–3593
    [Google Scholar]
  19. Ivins B. E., Wei kos S.L., Knudson G. B., Little S. F. 1990; Immunization against anthrax with aromatic compound-dependent (Aro-) mutants of Bacillus anthracis and with recombinant strains of Bacillus subtilis that produce anthrax protective antigen.. Infect Immun 58:303–308
    [Google Scholar]
  20. Leary S.E.C., Williamson E. D., Griffin K. F., Russell P., Eley S.M., Titball R. W. 1995; Active immunization with re-combinant V antigen from Yersinia pestis protects against plague. Infect Immun 63:2854–2858
    [Google Scholar]
  21. Levine M. M., Herrington D., Murphy J. R., Morris J. G., Losonsky G., Tall B., Lindberg A. A., Svenson S., Baqar S., Edwards M. F., Stocker B. 1987; Safety, infectivity, immuno- genicity and in vivo stability of two attenuated auxotrophic mutant strains of Salmonella typhi, 541 Ty and 543Ty, as live oral vaccines in humans.. J Clin Invest 79:888–902
    [Google Scholar]
  22. MacFarland W.C., Stocker B. A. D. 1987; Effect of different purine auxotrophic mutations on mouse-virulence of a Vi-positive strain of Salmonella dublin and two strains of Salmonella typhimurium.. Microb Pathog 3:129–141
    [Google Scholar]
  23. Marmur J. 1961; A procedure for the isolation of deoxyribonucleic acid from micro-organisms. J Mol Biol 3:208–218
    [Google Scholar]
  24. Marshall J. D., Bartelloni P. J., Cavanaugh D. C., Kadull P. J., Meyer K. F. 1974; Plague immunization. II. Relation of adverse clinical reactions to multiple immunizations with killed vaccine. J Infect Dis 129:S19–S25
    [Google Scholar]
  25. Maskell D., Sweeny K., O'Callaghan D., Hormaeche C., Liew F., Dougan G. 1987; Salmonella typhimurium aro A mutants as carriers of the Escherichia coli heat-labile enterotoxin B subunit to the secretory and systemic immune systems.. Microb Pathog 2:211–221
    [Google Scholar]
  26. McGhee J.R., Mestecky J., Dertzbaugh M. T., Eldridge J. H., Hirasava M., Kiyono H. 1992; The mucosal immune system: from fundamental concepts to vaccine development.. Vaccine 10:75–88
    [Google Scholar]
  27. Meyer K.F. 1970; Effectiveness of live or killed plague vaccines in man.. Bull WHO 42:653–666
    [Google Scholar]
  28. Meyer K. F., Cavanaugh D. C., Bartelloni P. J., Marshall J. D. 1974a; Plague immunization. I. Past and present trends. J Infect Dis S129:S13–S18
    [Google Scholar]
  29. Meyer K. F., Hightower J. A., McCrumb F.R. 1974b; Plague immunization. VI. Vaccination with the fraction 1 antigen of Yersinia pestis.. J Infect Dis S129:S41–S45
    [Google Scholar]
  30. Meyer K. F., Smith G., Foster L. E., Brookman M., Sung M. 1974c; Live, attenuated Yersinia pestis vaccine: virulent in nonhuman primates, harmless to guinea pigs.. J Infect Dis S129:S85–S120
    [Google Scholar]
  31. Miller V., Mekalanos J. 1988; A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires tox R. J Bacteriol 170:2575–2583
    [Google Scholar]
  32. Motin V. L., Nakajima R., Smirnov G. B., Brubaker R. R. 1994; Passive immunity to yersiniae mediated by anti-recombinant V antigen and protein A-V antigen fusion peptide. Infect Immun 62:4192–4201
    [Google Scholar]
  33. O'Gaora P., Maskell D., Coleman D., Cafferkey M., Dougan G. 1989; Cloning and characterization of the ser C and aro A genes of Yersinia enterocolitica and construction of an aro A mutant. Gene 84:23–30
    [Google Scholar]
  34. Oyston P.C.F., Williamson E. D., Leary S. E. C., Eley S. M., Griffin K. F., Titball R. W. 1995; Immunization with live recombinant Salmonella typhimurium aro A producing FI antigen protects against plague. Infect Immun 63:563–568
    [Google Scholar]
  35. Reed L.J, Si Muench H. 1938; A simple method for estimating fifty percent endpoints.. Am J Hyg 27:493–497
    [Google Scholar]
  36. Reisman R.E. 1907; Allergic reactions due to plague vaccine.. J Allergy 46:49–55
    [Google Scholar]
  37. Roberts M., Maskell D., Novotony P., Dougan G. 1990; Construction and characterization in vivo of Bordetella pertussis aro A mutants. Infect Immun 58:732–739
    [Google Scholar]
  38. Russell P., Eley S. M., Hibbs S. E., Manchee R. J., Stagg A. J., Titball R. W. 1995; A comparison of plague vaccine U.S.P. and EV76 vaccine induced protection against Yersinia pestis in a murine model.. Vaccine 13:1551–1556
    [Google Scholar]
  39. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2 nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  40. Sigwart D. F., Stocker B. A. D., Clements J. D. 1989; Effect of a pur A mutation on efficacy of Salmonella li ve-vaccine vectors. Infect Immun 57:1858–1861
    [Google Scholar]
  41. Straley S.C., Bowmer W. S. 1986; Virulence genes regulated at the transcriptional level by Ca2+ in Yersinia pestis include structural genes for outer membrane proteins. Infect Immun 51:445–454
    [Google Scholar]
  42. Straley S.C., Perry R. D. 1995; Environmental modulation of gene expression and pathogenesis in Yersinia. Trends Microbiol 3:310–317
    [Google Scholar]
  43. Vaughan L. M., Smith P. R., Foster T. J. 1993; An aromatic- dependent mutant of the fish pathogen Aeromonas salmonicida is attenuated in fish and is effective as a live vaccine against the salmonid disease furunculosis. Infect Immun 61:2172–2181
    [Google Scholar]
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