Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1986 Sep 1;164(3):868–881. doi: 10.1084/jem.164.3.868

Cloning of the structural genes of three H8 antigens and of protein III of Neisseria gonorrhoeae

PMCID: PMC2188392  PMID: 3091756

Abstract

A bank of gonococcal DNA was constructed in the lambda gt11 expression vector. immunological screening of the bank resulted in the isolation of a clone that contains the structural gene of protein III. In addition, several clones reactive with mAbs specific for the H8 antigen were isolated. DNA hybridization studies revealed that these H8- reactive clones were derived from three different gonococcal genes. When the products produced by these clones were used to absorb antibodies from a rabbit antiserum, and the eluted antibodies were used in immunological studies, it could be shown that the parent gonococcus expressed the product of two of these H8 genes, and in strain R10, these had Mr of approximately 19,700 21,200 respectively. The larger form has not been recognized hitherto because the epitope reactive with the H8 mAb may be masked in this product.

Full Text

The Full Text of this article is available as a PDF (1.5 MB).

Selected References

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

  1. Black W. J., Cannon J. G. Cloning of the gene for the common pathogenic Neisseria H.8 antigen from Neisseria gonorrhoeae. Infect Immun. 1985 Jan;47(1):322–325. doi: 10.1128/iai.47.1.322-325.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blake M. S., Gotschlich E. C. Purification and partial characterization of the major outer membrane protein of Neisseria gonorrhoeae. Infect Immun. 1982 Apr;36(1):277–283. doi: 10.1128/iai.36.1.277-283.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blake M. S., Gotschlich E. C. Purification and partial characterization of the opacity-associated proteins of Neisseria gonorrhoeae. J Exp Med. 1984 Feb 1;159(2):452–462. doi: 10.1084/jem.159.2.452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. doi: 10.1016/0003-2697(84)90320-8. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Cannon J. G., Buchanan T. M., Sparling P. F. Confirmation of association of protein I serotype of Neisseria gonorrhoeae with ability to cause disseminated infection. Infect Immun. 1983 May;40(2):816–819. doi: 10.1128/iai.40.2.816-819.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Diaz J. L., Heckels J. E. Antigenic variation of outer membrane protein II in colonial variants of Neisseria gonorrhoeae P9. J Gen Microbiol. 1982 Mar;128(3):585–591. doi: 10.1099/00221287-128-3-585. [DOI] [PubMed] [Google Scholar]
  9. Draper D. L., James J. F., Brooks G. F., Sweet R. L. Comparison of virulence markers of peritoneal and fallopian tube isolates with endocervical Neisseria gonorrhoeae isolates from women with acute salpingitis. Infect Immun. 1980 Mar;27(3):882–888. doi: 10.1128/iai.27.3.882-888.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hitchcock P. J., Hayes S. F., Mayer L. W., Shafer W. M., Tessier S. L. Analyses of gonococcal H8 antigen. Surface location, inter- and intrastrain electrophoretic heterogeneity, and unusual two-dimensional electrophoretic characteristics. J Exp Med. 1985 Dec 1;162(6):2017–2034. doi: 10.1084/jem.162.6.2017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hoopes B. C., McClure W. R. Studies on the selectivity of DNA precipitation by spermine. Nucleic Acids Res. 1981 Oct 24;9(20):5493–5504. doi: 10.1093/nar/9.20.5493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Johnston K. H., Gotschlich E. C. Isolation and characterization of the outer membrane of Neisseria gonorrhoeae. J Bacteriol. 1974 Jul;119(1):250–257. doi: 10.1128/jb.119.1.250-257.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Johnston K. H., Holmes K. K., Gotschlich E. C. The serological classification of Neisseria gonorrhoeae. I. Isolation of the outer membrane complex responsible for serotypic specificity. J Exp Med. 1976 Apr 1;143(4):741–758. doi: 10.1084/jem.143.4.741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Judd R. C. 125I-peptide mapping of protein III isolated from four strains of Neisseria gonorrhoeae. Infect Immun. 1982 Aug;37(2):622–631. doi: 10.1128/iai.37.2.622-631.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Judd R. C. Surface peptide mapping of protein I and protein III of four strains of Neisseria gonorrhoeae. Infect Immun. 1982 Aug;37(2):632–641. doi: 10.1128/iai.37.2.632-641.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Lambden P. R., Heckels J. E., James L. T., Watt P. J. Variations in surface protein composition associated with virulence properties in opacity types of Neisseria gonorrhoeae. J Gen Microbiol. 1979 Oct;114(2):305–312. doi: 10.1099/00221287-114-2-305. [DOI] [PubMed] [Google Scholar]
  18. McDade R. L., Jr, Johnston K. H. Characterization of serologically dominant outer membrane proteins of Neisseria gonorrhoeae. J Bacteriol. 1980 Mar;141(3):1183–1191. doi: 10.1128/jb.141.3.1183-1191.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Meinkoth J., Wahl G. Hybridization of nucleic acids immobilized on solid supports. Anal Biochem. 1984 May 1;138(2):267–284. doi: 10.1016/0003-2697(84)90808-x. [DOI] [PubMed] [Google Scholar]
  20. Nakamura K., Pirtle R. M., Inouye M. Homology of the gene coding for outer membrane lipoprotein within various Gram-negative bacteria. J Bacteriol. 1979 Jan;137(1):595–604. doi: 10.1128/jb.137.1.595-604.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Newhall W. J., Sawyer W. D., Haak R. A. Cross-linking analysis of the outer membrane proteins of Neisseria gonorrhoeae. Infect Immun. 1980 Jun;28(3):785–791. doi: 10.1128/iai.28.3.785-791.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sandstrom E. G., Knapp J. S., Buchanan T. B. Serology of Neisseria gonorrhoeae: W-antigen serogrouping by coagglutination and protein I serotyping by enzyme-linked immunosorbent assay both detect protein I antigens. Infect Immun. 1982 Jan;35(1):229–239. doi: 10.1128/iai.35.1.229-239.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schoolnik G. K., Buchanan T. M., Holmes K. K. Gonococci causing disseminated gonococcal infection are resistant to the bactericidal action of normal human sera. J Clin Invest. 1976 Nov;58(5):1163–1173. doi: 10.1172/JCI108569. [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. Swanson J., Barrera O. Immunological characteristics of gonococcal outer membrane protein II assessed by immunoprecipitation, immunoblotting, and coagglutination. J Exp Med. 1983 May 1;157(5):1405–1420. doi: 10.1084/jem.157.5.1405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Swanson J., Kraus S. J., Gotschlich E. C. Studies on gonococcus infection. I. Pili and zones of adhesion: their relation to gonococcal growth patterns. J Exp Med. 1971 Oct 1;134(4):886–906. doi: 10.1084/jem.134.4.886. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Swanson J. Studies on gonococcus infection. XII. Colony color and opacity varienats of gonococci. Infect Immun. 1978 Jan;19(1):320–331. doi: 10.1128/iai.19.1.320-331.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Swanson J. Studies on gonococcus infection. XIV. Cell wall protein differences among color/opacity colony variants of Neisseria gonorrhoeae. Infect Immun. 1978 Jul;21(1):292–302. doi: 10.1128/iai.21.1.292-302.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Weinberger C., Hollenberg S. M., Ong E. S., Harmon J. M., Brower S. T., Cidlowski J., Thompson E. B., Rosenfeld M. G., Evans R. M. Identification of human glucocorticoid receptor complementary DNA clones by epitope selection. Science. 1985 May 10;228(4700):740–742. doi: 10.1126/science.2581314. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES