Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1982 Dec;152(3):1241–1247. doi: 10.1128/jb.152.3.1241-1247.1982

Cloning of the chromosomal determinants encoding hemolysin production and mannose-resistant hemagglutination in Escherichia coli.

H Berger, J Hacker, A Juarez, C Hughes, W Goebel
PMCID: PMC221631  PMID: 6754701

Abstract

We have cloned the chromosomal hemolysin determinants from Escherichia coli strains belonging to the four O-serotypes O4, O6, O18, and O75. The hemolysin-producing clones were isolated from gene banks of these strains which were constructed by inserting partial Sau3A fragments of chromosomal DNA into the cosmid pJC74. The hemolytic cosmid clones were relatively stable. The inserts were further subcloned either as SalI fragments in pACYC184 or as BamHI-SalI fragments in a recombinant plasmid (pANN202) containing cistron C (hlyC) of the plasmid-encoded hemolysin determinant. Detailed restriction maps of each of these determinants were constructed, and it was found that, despite sharing overall homology, the determinants exhibited minor specific differences in their structure. These appeared to be restricted to cistron A (hlyA), which is the structural gene for hemolysin. In the gene banks of two of these hemolytic strains, we could also identify clones which carried the genetic determinants for the mannose-resistant hemagglutination antigens Vb and VIc. Both of these fimbrial antigens were expressed in the E. coli K-12 clones to an extent similar to that observed in the wild-type strains. These recombinant cosmids were rather unstable, and, in the absence of selection, segregated at a high frequency.

Full text

PDF
1242

Images in this article

1243Image
on p.1243
1244Image
on p.1244
1246Image
on p.1246

Selected References

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

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Collins J., Brüning H. J. Plasmids useable as gene-cloning vectors in an in vitro packaging by coliphage lambda: "cosmids". Gene. 1978 Oct;4(2):85–107. doi: 10.1016/0378-1119(78)90023-9. [DOI] [PubMed] [Google Scholar]
  4. Collins J., Hohn B. Cosmids: a type of plasmid gene-cloning vector that is packageable in vitro in bacteriophage lambda heads. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4242–4246. doi: 10.1073/pnas.75.9.4242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cooke E. M., Ewins S. P. Properties of strains of Escherichia coli isolated from a variety of sources. J Med Microbiol. 1975 Feb;8(1):107–111. doi: 10.1099/00222615-8-1-107. [DOI] [PubMed] [Google Scholar]
  6. DeBoy J. M., 2nd, Wachsmuth I. K., Davis B. R. Hemolytic activity in enterotoxigenic and non-enterotoxigenic strains of Escherichia coli. J Clin Microbiol. 1980 Aug;12(2):193–198. doi: 10.1128/jcm.12.2.193-198.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Evans D. J., Jr, Evans D. G., Höhne C., Noble M. A., Haldane E. V., Lior H., Young L. S. Hemolysin and K antigens in relation to serotype and hemagglutination type of Escherichia coli isolated from extraintestinal infections. J Clin Microbiol. 1981 Jan;13(1):171–178. doi: 10.1128/jcm.13.1.171-178.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Evans D. J., Jr, Evans D. G., Young L. S., Pitt J. Hemagglutination typing of Escherichia coli: definition of seven hemagglutination types. J Clin Microbiol. 1980 Aug;12(2):235–242. doi: 10.1128/jcm.12.2.235-242.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fried F. A., Vermeulen C. W., Ginsburg M. J., Cone C. M. Etiology of pyelonephritis: further evidence associating the production of experimental pyelonephritis with hemolysis in Escherichia coli. J Urol. 1971 Sep;106(3):351–354. doi: 10.1016/s0022-5347(17)61286-2. [DOI] [PubMed] [Google Scholar]
  10. Goebel W., Hedgpeth J. Cloning and functional characterization of the plasmid-encoded hemolysin determinant of Escherichia coli. J Bacteriol. 1982 Sep;151(3):1290–1298. doi: 10.1128/jb.151.3.1290-1298.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goebel W., Lindenmaier W., Pfeifer F., Schrempf H., Schelle B. Transposition and insertion of intact, deleted and enlarged ampicillin transposon Tn3 from mini-R1 (Rsc) plasmids into transfer factors. Mol Gen Genet. 1977 Nov 29;157(2):119–129. doi: 10.1007/BF00267389. [DOI] [PubMed] [Google Scholar]
  12. Goebel W., Royer-Pokora B., Lindenmaier W., Bujard H. Plasmids controlling synthesis of hemolysin in Escherichia coli: molecular properties. J Bacteriol. 1974 Jun;118(3):964–973. doi: 10.1128/jb.118.3.964-973.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hughes C., Phillips R., Roberts A. P. Serum resistance among Escherichia coli strains causing urinary tract infection in relation to O type and the carriage of hemolysin, colicin, and antibiotic resistance determinants. Infect Immun. 1982 Jan;35(1):270–275. doi: 10.1128/iai.35.1.270-275.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Minshew B. H., Jorgensen J., Counts G. W., Falkow S. Association of hemolysin production, hemagglutination of human erythrocytes, and virulence for chicken embryos of extraintestinal Escherichia coli isolates. Infect Immun. 1978 Apr;20(1):50–54. doi: 10.1128/iai.20.1.50-54.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Noegel A., Rdest U., Goebel W. Determination of the functions of hemolytic plasmid pHly152 of Escherichia coli. J Bacteriol. 1981 Jan;145(1):233–247. doi: 10.1128/jb.145.1.233-247.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Roberts A. P., Phillips R. Bacteria causing symptomatic urinary tract infection or asymptomatic bacteriuria. J Clin Pathol. 1979 May;32(5):492–496. doi: 10.1136/jcp.32.5.492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Smith H. W., Halls S. The transmissible nature of the genetic factor in Escherichia coli that controls haemolysin production. J Gen Microbiol. 1967 Apr;47(1):153–161. doi: 10.1099/00221287-47-1-153. [DOI] [PubMed] [Google Scholar]
  18. Springer W., Goebel W. Synthesis and secretion of hemolysin by Escherichia coli. J Bacteriol. 1980 Oct;144(1):53–59. doi: 10.1128/jb.144.1.53-59.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Waalwijk C., van den Bosch J. F., MacLaren D. M., de Graaff J. Hemolysin plasmid coding for the virulence of a nephropathogenic Escherichia coli strain. Infect Immun. 1982 Jan;35(1):32–37. doi: 10.1128/iai.35.1.32-37.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Welch R. A., Dellinger E. P., Minshew B., Falkow S. Haemolysin contributes to virulence of extra-intestinal E. coli infections. Nature. 1981 Dec 17;294(5842):665–667. doi: 10.1038/294665a0. [DOI] [PubMed] [Google Scholar]
  21. de la Cruz F., Müller D., Ortiz J. M., Goebel W. Hemolysis determinant common to Escherichia coli hemolytic plasmids of different incompatibility groups. J Bacteriol. 1980 Aug;143(2):825–833. doi: 10.1128/jb.143.2.825-833.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES