1887

Abstract

The gene coding for coagulase () was cloned from 8325-4 in a replacement vector in Coagulase (plasma-clotting) activity was measured in lysates and an immunoreactive protein of 60 kDa was detected by Western immunoblotting with anti-coagulase serum. This protein comigrated with the major immunoreactive protein in supernatants of 8325-4. The gene was subcloned in pUC vectors. One recombinant expressed a 60 kDa immunoreactive protein and plasma-clotting activity. A putative -galactosidase-coagulase fusion protein and truncated peptides were expressed by variants formed by subcloning. These results are consistent with previously published biochemical data that the prothrombin-binding domain of coagulase is located in the N-terminus of the protein. The cloned gene was transferred into on a shuttle plasmid. Expression of coagulase was higher in a strain with a mutation in the locus, which controls the level of several exoproteins in , suggesting that normally regulates coagulase expression negatively.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-134-1-75
1988-01-01
2021-08-05
Loading full text...

Full text loading...

/deliver/fulltext/micro/134/1/mic-134-1-75.html?itemId=/content/journal/micro/10.1099/00221287-134-1-75&mimeType=html&fmt=ahah

References

  1. Anderson J.C. 1983; Veterinary aspects of staphylococci. In Staphylococci and Staphylococcal Infections 1 pp. 193–237 Edited by Easmon C. S. F., Adlam C. London: Academic Press;
    [Google Scholar]
  2. Anderson J.C., Adlam C., Knights J.M. 1982; The effect of staphylocoagulase in the mammary gland of the mouse. British Journal of Experimental Pathology 63:336–340
    [Google Scholar]
  3. Appleyard R.K. 1954; Segregation of new lysogenic types during growth of a doubly lysogenic strain derived from Escherichia coli K12. Genetics 39:440–452
    [Google Scholar]
  4. Bjorklind A., Arvidson S. 1980; Mutants of Staphylococcus aureus affected in the regulation of exoprotein synthesis. FEMS Microbiology Letters 7:203–206
    [Google Scholar]
  5. Bolivar F., Rodriguez R.L., Greene P.J., Betlach M.V., Heynecker H.L., Boyer H.W., Crosa J.H., Falkow S. 1977; Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95–113
    [Google Scholar]
  6. Brown D.E., Pattee P.A. 1980; Identification of a chromosomal determinant of alpha-toxin production in Staphylococcus aureus. Infection and Immunity 30:36–42
    [Google Scholar]
  7. Burnette W.N. 1981; ‘Western blotting’: electrophoretic transfer of protein from sodium dodecyl sulphate polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Analytical Biochemistry 112:195–203
    [Google Scholar]
  8. Easmon C.S.F., Adlam C. 1983a Staphylococci and Staphylococcal Infections 1 London: Academic press;
    [Google Scholar]
  9. Easmon C.S.F., Adlam C. 1983b Staphylococci and Staphylococcal Infections 2 London: Academic press;
    [Google Scholar]
  10. Haraldsson I., Jonsson P. 1984; Histopathology and pathogenesis of mouse mastitis induced with Staphylococcus aureus. Journal of Comparative Pathology 94:183–196
    [Google Scholar]
  11. Hasegawa N., San Clemente C.L. 1978; Virulence and immunity of Staphylococcus aureus BB and certain deficient mutants. Infection and Immunity 22:473–479
    [Google Scholar]
  12. Helfman D.M., Fermamisco J.R., Fiddes J.C., Thomas G.P., Hughes S.H. 1983; Identification of clones that encode chicken tropomyosin by direct immunological screening of a cDNA expression library. Proceedings of the National Academy of Sciences of the United States of America 80:31–35
    [Google Scholar]
  13. Hemker H.C., Bas B.M., Muller A.D. 1975; Activation of a pro-enzyme by a stoichiometric reaction with another protein. The reaction between prothrombin and staphylocoagulase 379:180–188
    [Google Scholar]
  14. Jeljaszewicz J., Switalski L.M., Adlam C. 1983; Staphylocoagulase and clumping factor. In Staphylococci and Staphylococcal Infections 2 pp. 525–557 Edited by Easmon C. S. F., Adlam C. London: Academic Press;
    [Google Scholar]
  15. Jones C.L., Khan S.A. 1986; Nucleotide sequence of the enterotoxin B gene from Staphylococcus aureus. Journal of Bacteriology 166:29–33
    [Google Scholar]
  16. Jonsson P., Lindberg M., Haraldsson I., Wadstrom T. 1985; Virulence of Staphylococcus aureus in a mouse mastitis model: studies of alpha hemolysin, coagulase, and protein A as possible virulence determinants with protoplast fusion and gene cloning. Infection and Immunity 49:765–769
    [Google Scholar]
  17. Kawabata S., Miyata T., Morita T., Miyata T., Iwanaga S., Igarishi H. 1986a; The amino acid sequence of the procoagulant- and prothrombinbinding domain isolated from staphylocoagulase. Journal of Biological Chemistry 261:527–531
    [Google Scholar]
  18. Kawabata S., Morita T., Miyata T., Iwanaga S., Igarishi H. 1986b; Isolation and characterization of staphylocoagulase chymotryptic fragment. Journal of Biological Chemistry 261:1427–1433
    [Google Scholar]
  19. Kinsman O.S., Arbuthnott J.P. 1980; Experimental staphylococcal infections in newborn mice: inhibition of weight gain as an index of virulence. Journal of Medical Microbiology 13:281–290
    [Google Scholar]
  20. Kreiswirth B.N., Lofdahl S., Betley M.J., O’Reilly M., Schlievert P.M., Bergdoll M.S., Novick R.P. 1983; The toxic shock syndrome exotoxin structural gene is not detectably transmitted by a prophage. Nature, London 305:709–712
    [Google Scholar]
  21. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  22. Loenen W.A.M., Brammar W.J. 1980; A bacteriophage vector for cloning large restriction fragments made with several restriction enzymes. Gene 10:249–259
    [Google Scholar]
  23. Lofdahl S., Guss B., Uhlen M., Philipson L., Lindberg M. 1983; Gene for staphylococcal protein A. Proceedings of the National Academy of Sciences of the United States of America 80:697–701
    [Google Scholar]
  24. Mccartney C., Arbuthnott J.P. 1978; Mode of action of membrane-damaging toxins produced by staphylococci. In Bacterial Toxins and Cell Membranes pp. 89–122 Edited by Jeljaszewicz J., Wadstrom T. London: Academic Press;
    [Google Scholar]
  25. Maniatis T., Fritsch E.F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  26. Masuda S. 1983; An efficient method for the isolation of a mutant with extremely low producibility of coagulase from a Staphylococcus aureus strain. Microbiology and Immunology 27:801–805
    [Google Scholar]
  27. Miller J.H. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  28. Novick R.P. 1963; Properties of a cryptic high- frequency transducing phage in Staphylococcus aureus. Virology 33:155–166
    [Google Scholar]
  29. O’Reilly M., De Azavedo J.C.S., Kennedy S., Foster T.J. 1986; Inactivation of the alpha- haemolysin of Staphylococcus aureus 8325-4 by site- directed mutagenesis and studies on expression of its haemolysins. Microbial Pathogenesis 1:125–138
    [Google Scholar]
  30. O’Toole P.W., Foster T.J. 1986a; Molecular cloning and expression of the epidermolytic toxin A gene of Staphylococcus aureus. Microbial Pathogenesis 1:583–594
    [Google Scholar]
  31. O’Toole P.W., Foster T.J. 1986b; Epidermolytic toxin serotype B of Staphylococcus aureus is plasmid-encoded. FEMS Microbiology Letters 36:311–314
    [Google Scholar]
  32. Patel A.H., Nowlan P., Weavers E.D., Foster T.J. 1987; Studies on the virulence of protein-A- deficient and a-toxin-deficient mutants of Staphylococcus aureus isolated by allele replacement. Infection and Immunity 55: (in the Press)
    [Google Scholar]
  33. Ranneli D.M., Jones C.L., Johns M.B., Mussey G.J., Khan S.A. 1985; Molecular cloning of staphylococcal enterotoxin B gene in Escherichia coli and Staphylococcus aureus. Proceedings of the National Academy of Sciences of the United States of America 82:5850–5854
    [Google Scholar]
  34. Recsei P., Kreiswirth B., O’Reilly M., Schlievert P., Gruss A., Novick R.P. 1986; Regulation of exoprotein gene expression in Staphylococcus aureus by agr. Molecular and General Genetics 202:58–61
    [Google Scholar]
  35. Russell R.R.B., Coleman D., Dougan G. 1985; Expression of a gene for glucan-binding protein from Streptococcus mutans in Escherichia coli. Journal of General Microbiology 131:295–299
    [Google Scholar]
  36. Smith D.D., Johnstone J.M. 1958; Staphylocoagulase activity in vivo. British Journal of Experimental Pathology 39:165–170
    [Google Scholar]
  37. Uhlen M., Guss B., Nilsson B., Gotz F., Lindberg M. 1984; Expression of the gene encoding protein A in Staphylococcus aureus and coagulase-negative staphylococci. Journal of Bacteriology 159:713–719
    [Google Scholar]
  38. Yanish-Perron C., Vieira J.C., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequehces of M13mpl8 and pUC19 vectors. Gene 33:103–119
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-134-1-75
Loading
/content/journal/micro/10.1099/00221287-134-1-75
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error