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. 1988 Sep;85(18):6841–6845. doi: 10.1073/pnas.85.18.6841

Genetics of protein I of Neisseria gonorrhoeae: construction of hybrid porins.

N H Carbonetti 1, V I Simnad 1, H S Seifert 1, M So 1, P F Sparling 1
PMCID: PMC282074  PMID: 2457919

Abstract

Protein I (PI), the major outer membrane protein of Neisseria gonorrhoeae, is a porin and occurs in two major immunochemical classes, A and B. By using shuttle mutagenesis to insert a selectable marker close to the PI structural gene, evidence was obtained from transformation experiments to demonstrate that the PI structural gene is equivalent to the defined locus nmp and that the genes for PI class A and PI class B are alleles of the same locus. The PI class B gene of strain MS11 was cloned and sequenced, and comparison of this sequence with the gene sequence of PI class A of FA19 revealed a number of regions of significant divergence. By selection for the closely linked marker in transformations between the two strains, a series of strains with a hybrid PI was obtained. Analysis of these strains with monoclonal antibodies and oligonucleotides specific to PI class A or PI class B elucidated the nature and location of some of the surface-exposed epitopes, a thorough characterization of which is a prerequisite for understanding the role of PI in gonococcal pathogenesis and its possible use as a component of a vaccine.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Biswas G. D., Sox T., Blackman E., Sparling P. F. Factors affecting genetic transformation of Neisseria gonorrhoeae. J Bacteriol. 1977 Feb;129(2):983–992. doi: 10.1128/jb.129.2.983-992.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blake M. S., Gotschlich E. C., Swanson J. Effects of proteolytic enzymes on the outer membrane proteins of Neisseria gonorrhoeae. Infect Immun. 1981 Jul;33(1):212–222. doi: 10.1128/iai.33.1.212-222.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buchanan T. M., Hildebrandt J. F. Antigen-specific serotyping of Neisseria gonorrhoeae: characterization based upon principal outer membrane protein. Infect Immun. 1981 Jun;32(3):985–994. doi: 10.1128/iai.32.3.985-994.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cannon J. G., Black W. J., Nachamkin I., Stewart P. W. Monoclonal antibody that recognizes an outer membrane antigen common to the pathogenic Neisseria species but not to most nonpathogenic Neisseria species. Infect Immun. 1984 Mar;43(3):994–999. doi: 10.1128/iai.43.3.994-999.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cannon J. G., Klapper D. G., Blackman E. Y., Sparling P. F. Genetic locus (nmp-1) affecting the principal outer membrane protein of Neisseria gonorrhoeae. J Bacteriol. 1980 Aug;143(2):847–851. doi: 10.1128/jb.143.2.847-851.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cannon J. G., Lee T. J., Guymon L. F., Sparling P. F. Genetics of serum resistance in Neisseria gonorrhoeae: the sac-1 genetic locus. Infect Immun. 1981 May;32(2):547–552. doi: 10.1128/iai.32.2.547-552.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Carbonetti N. H., Sparling P. F. Molecular cloning and characterization of the structural gene for protein I, the major outer membrane protein of Neisseria gonorrhoeae. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9084–9088. doi: 10.1073/pnas.84.24.9084. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Danielsson D., Faruki H., Dyer D., Sparling P. F. Recombination near the antibiotic resistance locus penB results in antigenic variation of gonococcal outer membrane protein I. Infect Immun. 1986 May;52(2):529–533. doi: 10.1128/iai.52.2.529-533.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gotschlich E. C., Seiff M. E., Blake M. S., Koomey M. Porin protein of Neisseria gonorrhoeae: cloning and gene structure. Proc Natl Acad Sci U S A. 1987 Nov;84(22):8135–8139. doi: 10.1073/pnas.84.22.8135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Joiner K. A., Warren K. A., Tam M., Frank M. M. Monoclonal antibodies directed against gonococcal protein I vary in bactericidal activity. J Immunol. 1985 May;134(5):3411–3419. [PubMed] [Google Scholar]
  11. Judd R. C. Evidence for N-terminal exposure of the protein IA subclass of Neisseria gonorrhoeae protein I. Infect Immun. 1986 Nov;54(2):408–414. doi: 10.1128/iai.54.2.408-414.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. KELLOGG D. S., Jr, PEACOCK W. L., Jr, DEACON W. E., BROWN L., PIRKLE D. I. NEISSERIA GONORRHOEAE. I. VIRULENCE GENETICALLY LINKED TO CLONAL VARIATION. J Bacteriol. 1963 Jun;85:1274–1279. doi: 10.1128/jb.85.6.1274-1279.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Knapp J. S., Tam M. R., Nowinski R. C., Holmes K. K., Sandström E. G. Serological classification of Neisseria gonorrhoeae with use of monoclonal antibodies to gonococcal outer membrane protein I. J Infect Dis. 1984 Jul;150(1):44–48. doi: 10.1093/infdis/150.1.44. [DOI] [PubMed] [Google Scholar]
  14. Lambden P. R., Robertson J. N., Watt P. J. Biological properties of two distinct pilus types produced by isogenic variants of Neisseria gonorrhoeae P9. J Bacteriol. 1980 Jan;141(1):393–396. doi: 10.1128/jb.141.1.393-396.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lynch E. C., Blake M. S., Gotschlich E. C., Mauro A. Studies of Porins: Spontaneously Transferred from Whole Cells and Reconstituted from Purified Proteins of Neisseria gonorrhoeae and Neisseria meningitidis. Biophys J. 1984 Jan;45(1):104–107. doi: 10.1016/S0006-3495(84)84127-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Maness M. J., Sparling P. F. Multiple antibiotic resistance due to a single mutation in Neisseria gonorrhoeae. J Infect Dis. 1973 Sep;128(3):321–330. doi: 10.1093/infdis/128.3.321. [DOI] [PubMed] [Google Scholar]
  17. Meyer T. F., Mlawer N., So M. Pilus expression in Neisseria gonorrhoeae involves chromosomal rearrangement. Cell. 1982 Aug;30(1):45–52. doi: 10.1016/0092-8674(82)90010-1. [DOI] [PubMed] [Google Scholar]
  18. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  19. Sandstrom E. G., Chen K. C., Buchanan T. M. Serology of Neisseria gonorrhoeae: coagglutination serogroups WI and WII/III correspond to different outer membrane protein I molecules. Infect Immun. 1982 Nov;38(2):462–470. doi: 10.1128/iai.38.2.462-470.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sarubbi F. A., Jr, Blackman E., Sparling P. F. Genetic mapping of linked antibiotic resistance loci in Neisseria gonorrhoeae. J Bacteriol. 1974 Dec;120(3):1284–1292. doi: 10.1128/jb.120.3.1284-1292.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schneider H., Hale T. L., Zollinger W. D., Seid R. C., Jr, Hammack C. A., Griffiss J. M. Heterogeneity of molecular size and antigenic expression within lipooligosaccharides of individual strains of Neisseria gonorrhoeae and Neisseria meningitidis. Infect Immun. 1984 Sep;45(3):544–549. doi: 10.1128/iai.45.3.544-549.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schwalbe R. S., Sparling P. F., Cannon J. G. Variation of Neisseria gonorrhoeae protein II among isolates from an outbreak caused by a single gonococcal strain. Infect Immun. 1985 Jul;49(1):250–252. doi: 10.1128/iai.49.1.250-252.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Seifert H. S., Chen E. Y., So M., Heffron F. Shuttle mutagenesis: a method of transposon mutagenesis for Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1986 Feb;83(3):735–739. doi: 10.1073/pnas.83.3.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stern A., Nickel P., Meyer T. F., So M. Opacity determinants of Neisseria gonorrhoeae: gene expression and chromosomal linkage to the gonococcal pilus gene. Cell. 1984 Jun;37(2):447–456. doi: 10.1016/0092-8674(84)90375-1. [DOI] [PubMed] [Google Scholar]
  25. Teerlink T., Versantvoort H., Beuvery E. C. Antigenic and immunogenic properties of cyanogen bromide peptides from gonococcal outer membrane protein IB. Evidence for the existence of a surface-exposed conserved epitope. J Exp Med. 1987 Jul 1;166(1):63–76. doi: 10.1084/jem.166.1.63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Virji M., Fletcher J. N., Zak K., Heckels J. E. The potential protective effect of monoclonal antibodies to gonococcal outer membrane protein IA. J Gen Microbiol. 1987 Sep;133(9):2639–2646. doi: 10.1099/00221287-133-9-2639. [DOI] [PubMed] [Google Scholar]
  27. Young J. D., Blake M., Mauro A., Cohn Z. A. Properties of the major outer membrane protein from Neisseria gonorrhoeae incorporated into model lipid membranes. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3831–3835. doi: 10.1073/pnas.80.12.3831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Young R. A., Davis R. W. Yeast RNA polymerase II genes: isolation with antibody probes. Science. 1983 Nov 18;222(4625):778–782. doi: 10.1126/science.6356359. [DOI] [PubMed] [Google Scholar]
  29. Zak K., Diaz J. L., Jackson D., Heckels J. E. Antigenic variation during infection with Neisseria gonorrhoeae: detection of antibodies to surface proteins in sera of patients with gonorrhea. J Infect Dis. 1984 Feb;149(2):166–174. doi: 10.1093/infdis/149.2.166. [DOI] [PubMed] [Google Scholar]

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