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. 1982 Nov;38(2):739–744. doi: 10.1128/iai.38.2.739-744.1982

Cloning of genes determining the production of mannose-resistant fimbriae in a uropathogenic strain of Escherichia coli belonging to serogroup O6.

S Clegg
PMCID: PMC347800  PMID: 6128310

Abstract

Chromosomal DNA from a uropathogenic strain of Escherichia coli was partially digested with the restriction enzyme EcoRI. The partial digests were ligated into a cosmid containing an ampicillin-resistant determinant and packaged into lambda phage particles. An ampicillin-resistant transductant of E. coli HB101 was found to possess mannose-resistant hemagglutinating activity associated with a 50-kilobase-pair plasmid. Subcloning of the mannose-resistant fimbrial genes revealed that the genetic determinants were encoded by a 6.9-kilobase-pair DNA fragment of a recombinant plasmid. Chimeric plasmids smaller in size were unable to transform E. coli to fimbrial production. Physical maps of the recombinant plasmids were prepared showing restriction endonuclease sites within the inserted DNA fragments. The hemagglutinating activities of the wild-type strain and of the recombinant derivative were compared. Both strains agglutinated human erythrocytes in the presence of D-mannose to the same degree and also failed to produce fimbriae after incubation at 18 degrees C. Also, both strains were agglutinated by antifimbrial serum at a high titer, whereas no such activity was observed when a strain of E. coli which did not possess a plasmid was used.

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Selected References

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  1. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  2. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  3. Cameron J. R., Philippsen P., Davis R. W. Analysis of chromosomal integration and deletions of yeast plasmids. Nucleic Acids Res. 1977;4(5):1429–1448. doi: 10.1093/nar/4.5.1429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clegg S. Serological heterogeneity among fimbrial antigens causing mannose-resistant hemagglutination by uropathogenic Escherichia coli. Infect Immun. 1982 Feb;35(2):745–748. doi: 10.1128/iai.35.2.745-748.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clewell D. B. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol. J Bacteriol. 1972 May;110(2):667–676. doi: 10.1128/jb.110.2.667-676.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Duguid J. P., Clegg S., Wilson M. I. The fimbrial and non-fimbrial haemagglutinins of Escherichia coli. J Med Microbiol. 1979 May;12(2):213–227. doi: 10.1099/00222615-12-2-213. [DOI] [PubMed] [Google Scholar]
  8. Evans D. G., Evans D. J., Jr, Clegg S., Pauley J. A. Purification and characterization of the CFA/I antigen of enterotoxigenic Escherichia coli. Infect Immun. 1979 Aug;25(2):738–748. doi: 10.1128/iai.25.2.738-748.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Evans D. G., Evans D. J., Jr New surface-associated heat-labile colonization factor antigen (CFA/II) produced by enterotoxigenic Escherichia coli of serogroups O6 and O8. Infect Immun. 1978 Aug;21(2):638–647. doi: 10.1128/iai.21.2.638-647.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Evans D. G., Silver R. P., Evans D. J., Jr, Chase D. G., Gorbach S. L. Plasmid-controlled colonization factor associated with virulence in Esherichia coli enterotoxigenic for humans. Infect Immun. 1975 Sep;12(3):656–667. doi: 10.1128/iai.12.3.656-667.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Green C. P., Thomas V. L. Hemagglutination of human type O erythrocytes, hemolysin production, and serogrouping of Escherichia coli isolates from patients with acute pyelonephritis, cystitis, and asymptomatic bacteriuria. Infect Immun. 1981 Jan;31(1):309–315. doi: 10.1128/iai.31.1.309-315.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Guerry P., LeBlanc D. J., Falkow S. General method for the isolation of plasmid deoxyribonucleic acid. J Bacteriol. 1973 Nov;116(2):1064–1066. doi: 10.1128/jb.116.2.1064-1066.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hull R. A., Gill R. E., Hsu P., Minshew B. H., Falkow S. Construction and expression of recombinant plasmids encoding type 1 or D-mannose-resistant pili from a urinary tract infection Escherichia coli isolate. Infect Immun. 1981 Sep;33(3):933–938. doi: 10.1128/iai.33.3.933-938.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jones G. W., Rutter J. M. The association of K88 antigen with haemagglutinating activity in porcine strains of Escherichia coli. J Gen Microbiol. 1974 Sep;84(1):135–144. doi: 10.1099/00221287-84-1-135. [DOI] [PubMed] [Google Scholar]
  15. Kehoe M., Sellwood R., Shipley P., Dougan G. Genetic analysis of K88-mediated adhesion of enterotoxigenic Escherichia coli. Nature. 1981 May 14;291(5811):122–126. doi: 10.1038/291122a0. [DOI] [PubMed] [Google Scholar]
  16. Källenius G., Möllby R., Winberg J. In vitro adhesion of uropathogenic Escherichia coli to human periurethral cells. Infect Immun. 1980 Jun;28(3):972–980. doi: 10.1128/iai.28.3.972-980.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ljungh A., Faris A., Wadström T. Hemagglutination by Escherichia coli in septicemia and urinary tract infections. J Clin Microbiol. 1979 Oct;10(4):477–481. doi: 10.1128/jcm.10.4.477-481.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Meyerowitz E. M., Guild G. M., Prestidge L. S., Hogness D. S. A new high-capacity cosmid vector and its use. Gene. 1980 Nov;11(3-4):271–282. doi: 10.1016/0378-1119(80)90067-0. [DOI] [PubMed] [Google Scholar]
  19. Morrison D. A. Transformation in Escherichia coli: cryogenic preservation of competent cells. J Bacteriol. 1977 Oct;132(1):349–351. doi: 10.1128/jb.132.1.349-351.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Orskov I., Orskov F., Birch-Andersen A. Comparison of Escherichia coli fimbrial antigen F7 with type 1 fimbriae. Infect Immun. 1980 Feb;27(2):657–666. doi: 10.1128/iai.27.2.657-666.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Orskov I., Orskov F., Smith H. W., Sojka W. J. The establishment of K99, a thermolabile, transmissible escherichia coli K antigen, previously called "Kco", possessed by calf and lamb enteropathogenic strains. Acta Pathol Microbiol Scand B. 1975 Feb;83(1):31–36. doi: 10.1111/j.1699-0463.1975.tb00066.x. [DOI] [PubMed] [Google Scholar]
  22. Selker E., Brown K., Yanofsky C. Mitomycin C-induced expression of trpA of Salmonella typhimurium inserted into the plasmid ColE1. J Bacteriol. 1977 Jan;129(1):388–394. doi: 10.1128/jb.129.1.388-394.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Stauffer G. V., Plamann M. D., Stauffer L. T. Construction and expression of hybrid plasmids containing the Escherichia coli glyA genes. Gene. 1981 Jun-Jul;14(1-2):63–72. doi: 10.1016/0378-1119(81)90148-7. [DOI] [PubMed] [Google Scholar]
  24. Williams J. A., Yeggy J. P., Field C. C., Markovetz A. J. Resistance plasmids in Pseudomonas cepacia 4G9. J Bacteriol. 1979 Dec;140(3):1017–1022. doi: 10.1128/jb.140.3.1017-1022.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]

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