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Colonization factor antigen II (CFA/II) of enterotoxigenic Escherichia coli: molecular cloning of the CS3 determinant

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Summary

The genes for the cell surface associated antigen CS3, produced by CFA/II type enterotoxigenic Escherichia coli, have been cloned in the plasmid vector pBR322 to produce a family of recombinant plasmids. These plasmids contain a series of HindIII fragments of which a fragment of 4.6 kb is common to all those expressing CS3. One of these plasmids, pPM474, has been subjected to mutagenesis with Tn1725 and deletions generated using Bal31. This has defined a minimum region of 3.75 kb necessary for the production of CS3 on the cell surface and implying genetic complexity as has been observed with other fimbrial antigens. Analysis of the plasmid encoded proteins in E. coli K-12 minicells has confirmed this complexity.

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Abbreviations

Ap:

ampicillin

kb:

kilobase pairs

R (superscript):

resistant

S (superscript):

sensitive

Tc:

tetracycline

References

  • Achtman M, Schwuchow S, Helmuth R, Morelli G, Manning PA (1978) Cell-cell interactions in conjugating Escherichia coli: Con− mutants and stabilization of mating aggregates. Mol Gen Genet 164:171–183

    Google Scholar 

  • Achtman M, Manning PA, Edelbluth C, Herrlich P (1979) Export without proteolytic processing of inner and outer membrane proteins encoded by F sex factor tra cistrons in Escherichia coli minicells. Proc Natl Acad Sci USA 76:4837–4841

    Google Scholar 

  • Altenbuchner J, Schmid K, Schmitt R (1983) Tn1721-encoded tetracycline resistance: mapping of structural and regulatory genes mediating resistance. J Bacteriol 153:116–123

    Google Scholar 

  • Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heynecker HL, Boyer HW, Crosa JH, Falkow S (1977) Construction and characterization of new cloning vectors II a multipurpose cloning system. Gene 2:95–113

    Google Scholar 

  • Brown MCM, Weston A, Saunders JR, Humphreys GO (1979) Transformation of E. coli C600 by plasmid DNA at different phases of growth. FEMS Microbiol Lett 5:219–222

    Google Scholar 

  • Clewell DB, Helinski DR (1968) Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an open circular DNA form. Proc Natl Acad Sci USA 62:1159–1166

    Google Scholar 

  • Clewell DB, Helinski DR (1970) Properties of supercoiled deoxyribonucleic acid-protein relaxation complex and strand specificity of the relaxation event. Biochemistry 9:4428–4440

    Google Scholar 

  • Cravioto A, Scotland SM, Rowe B (1982) Haemagglutination activity and colonization factor antigens I and II in enterotoxigenic and non-enterotoxigenic strains of Escherichia coli isolated from humans. Infect Immun 36:189–197

    Google Scholar 

  • Darfeuille A, Lafeuille B, Joly B, Cluzel R (1983) A new colonization factor antigen (CFA/III) produced by enterotoxigenic Escherichia coli O128:B12. Ann Microbiol Inst Pasteur 134A:53–64

    Google Scholar 

  • Evans DG, Evans DJ (1978) New surface associated heat-labile colonization factor antigen (CFA/II) produced by enterotoxigenic Escherichia coli of serogroups 06 and 08. Infect Immun 21:638–647

    Google Scholar 

  • Evans DG, Evans DJ Jr, Tjoa W (1977) Haemagglutination of human group A erythrocytes by enterotoxigenic and enteropathogenic Escherichia coli isolated from adults with diarrhoea: correlation with colonization factor. Infect Immun 18:330–337

    Google Scholar 

  • Evans DG, Silver RP, Evans DJ Jr, Chase DG, Gorbach SL (1975) Plasmid-controlled colonization factor associated with virulence in Escherichia coli enterotoxigenic for humans. Infect Immun 12:656–667

    Google Scholar 

  • Evans DG, Evans DJ, Tjoa WS, Du Pont HL (1978) Detection and characterization of colonization factors of enterotoxigenic Escherichia coli isolated from adults with diarrhoea. Infect Immun 19:727–736

    Google Scholar 

  • Gaastra W, de Graaf FK (1982) Host-specific fimbrial adhesins of noninvasive enterotoxigenic Escherichia coli strains. Microbiol Rev 46:129–161

    Google Scholar 

  • Hawkes R, Niday E, Gordon J (1982) A dot-immuno-binding assay for monoclonal and other antibodies. Anal Biochem 119:142–147

    Google Scholar 

  • Henning U, Schwarz H, Chen R (1979) Radioimmunological screening method for specific membrane proteins. Anal Biochem 97:153–157

    Google Scholar 

  • Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–195

    Google Scholar 

  • Honda T, Arita M, Miwatani T (1984) Characterization of new hydrophobic pili of human enterotoxigenic Escherichia coli: a possible new colonization factor. Infect Immun 43:959–965

    Google Scholar 

  • Humphreys GO, Willshaw GA, Anderson ES (1975) A simple method for the preparation of large quantities of pure plasmid DNA. Biochim Biophys Acta 383:457–463

    Google Scholar 

  • Kado CI, Liu ST (1978) Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol 145:1365–1373

    Google Scholar 

  • Kennedy N, Beutin L, Achtman M, Skurray R, Rahmsdorf U, Herrlich P (1977) Conjugation proteins of the F sex factor. Nature 270:580–585

    Google Scholar 

  • Levine MM, Ristaino P, Marley G, Smyth C, Knutton S, Boedeker E, Black R, Young C, Clements ML, Cheney C, Patnaik R (1984) Coli surface antigens 1 and 3 of colonization factor antigen II — positive enterotoxigenic Escherichia coli: morphology, purification and immune responses in humans. Infect Immun 44:409–420

    Google Scholar 

  • Lugtenberg B, Meijers J, Peters R, Van der Hoek P, Van Alphen L (1975) Electrophoretic resolution of the ‘major outer membrane protein’ of Escherichia coli K-12 into four bands. FEBS Lett 58:254–258

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, New York

    Google Scholar 

  • Manning PA, Brown MH, Heuzenroeder MW (1984) Cloning of the structural gene (hly) for the haemolysin of Vibrio cholerae El Tor strain O17. Gene 31:235–241

    Google Scholar 

  • Moore D, Sowa BA, Ippen-Ihler K (1981) Location of an F-pilin pool in the inner membrane. J Bacteriol 146:251–259

    Google Scholar 

  • Mullany P, Field AM, McConnell MM, Scotland SM, Smith HR, Rowe B (1983) Expression of plasmids coding for colonization factor antigen II (CFA/II) and enterotoxin production in Escherichia coli. J Gen Microbiol 129:3591–3601

    Google Scholar 

  • Smyth CJ (1982) Two mannose-resistant haemagglutinins on enterotoxigenic Escherichia coli of serotype O6:K15:H16 or H− isolated from travellers' and infantile diarrhoea. J Gen Microbiol 128:2081–2096

    Google Scholar 

  • Stirm S, Orskov I, Orskov F, Mausa B (1967) Episome-carried surface antigen K88 of Escherichia coli. II. Isolation and chemical analysis. J Bacteriol 93:731–739

    Google Scholar 

  • Thomas LV, Cravioto A, Scotland SM, Rowe B (1982) New fimbrial antigen type (E8775) that may represent a new colonization factor in enterotoxigenic Escherichia coli in humans. Infect Immun 35:1119–1124

    Google Scholar 

  • Towbin H, Staehlin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

    Google Scholar 

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Author notes

  1. Kenneth N. Timmis

    Present address: Department of Medical Biochemistry, University Medical Centre, University of Geneva, Switzerland

Authors and Affiliations

  1. Department of Microbiology and Immunology, The University of Adelaide, 5001, Adelaide, S.A., Australia

    Paul A. Manning, Kenneth N. Timmis & Gordon Stevenson

Authors
  1. Paul A. Manning
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  2. Kenneth N. Timmis
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  3. Gordon Stevenson
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Communicated by E. Bautz

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Manning, P.A., Timmis, K.N. & Stevenson, G. Colonization factor antigen II (CFA/II) of enterotoxigenic Escherichia coli: molecular cloning of the CS3 determinant. Molec. Gen. Genet. 200, 322–327 (1985). https://doi.org/10.1007/BF00425443

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  • DOI: https://doi.org/10.1007/BF00425443

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