1887

Abstract

A novel klebicin, klebicin B, produced by an isolate of has been identified. It is encoded by a 5.5 kb plasmid, pKlebB-K 17/80, which is mobilized into UNF5023 by a large plasmid found in the same strain. The 5.5 kb plasmid has been cloned into the high-copy-number vector pUC19 and the restriction map of the resulting recombinant plasmid pRJ180 has been determined. Using sub-cloning and transposon mutagenesis, the klebicin B structural gene, the klebicin B immunity gene and the mitomycin C (MC) sensitivity gene () present on pRJ180 have been localized. Transposon inserts which inactivated klebicin production also abolished lysis protein production encoded by pRJ180, but did not affect klebicin B immunity. Using SDS-PAGE an MC-induced polypeptide of 85 kDa was observed in cultures of UNF5023(pRJ180). This polypeptide was absent in cultures carrying plasmid pRJ180 with a Tn insert which inactivated klebicin production. Analysis of the polypeptides present in the medium of JM83 (pRJ180) or UNF5023(pRJ180) indicated that the 85 kDa polypeptide is specifically secreted from the producing cell. Klebicin B has been purified, using gel filtration, from a cell-free extract of UNF5023(pRJ180) which had been induced with MC. After boiling in sample buffer the purified klebicin B gave rise to two peptides on SDS-PAGE, one of 85 kDa and the other of 11 kDa. Klebicin B-resistant mutants of UNF5023 were sensitive to klebicin A, colicin B and colicin D.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-134-9-2525
1988-09-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/134/9/mic-134-9-2525.html?itemId=/content/journal/micro/10.1099/00221287-134-9-2525&mimeType=html&fmt=ahah

References

  1. Bauernfeind A., Petermuller C., Schneider R. 1981; Bacteriocins as tools in analysis of nosocomial Klebsiella pneumoniae infections.. Journal of Clinical Microbiology 14:15–19
    [Google Scholar]
  2. Birnboim H.C., Doly J. 1979; A rapid alkaline extraction procedure for screening plasmid DNA.. Nucleic Acids Research 7:1513–1523
    [Google Scholar]
  3. Buffenmeyer C.L., Rychek R.R., Yee R.B. 1976; Bacteriocin (klebocin) sensitivity typing of Klebsiella.. Journal of Clinical Microbiology 4:239–244
    [Google Scholar]
  4. Chak K.-F., James R. 1984; Localization and characterization of a gene on the ColE3-CA38 plasmid that confers immunity to colicin E8.. Journal of General Microbiology 130:701–710
    [Google Scholar]
  5. Chak K.-F., James R. 1985; Analysis of the promoters for the two immunity genes present in the ColE3-CA38 plasmid using two new promoter probe vectors.. Nucleic Acids Research 13:2519–2531
    [Google Scholar]
  6. Chak K.-F., James R. 1986; Characterization of the ColE9-J plasmid and analysis of its genetic organization.. Journal of General Microbiology 132:61–71
    [Google Scholar]
  7. Cooper P.C., James R. 1985; Three immunity types of klebicins which use the cloacin DF13 receptor of Klebsiella pneumoniae.. Journal of General Microbiology 131:2313–2318
    [Google Scholar]
  8. Cross A., Allen J.R., Burke J., Ducel G., Harris A., John J., Johnson D., Lew M., Macmillan B., Meers P., Skalove R., Wenzel R., Tenney J. 1983; Nosocomial infections due to Pseudomonas aeruginosa: review of recent trends.. Reviews of Infectious Diseases 5: Supplement 5 S837–S845
    [Google Scholar]
  9. Cryz S.J., Mortimer P.M., Mansfield V., Germanier R. 1986; Seroepidemiology of Klebsiellabacteremic isolates and implications for vaccine development.. Journal of Clinical Microbiology 23:687–690
    [Google Scholar]
  10. Edmondson A.S., Cooke E.M. 1979; The development and assessment of a bacteriocin typing method for Klebsiella.. Journal of Hygiene 82:207–223
    [Google Scholar]
  11. Gaston M.A., Ayling-Smith B., Pitt T.Y. 1987; New bacteriophage typing scheme for subdivision of the frequent capsular serotypes of Klebsiellaspp.. Journal of Clinical Microbiology 25:1228–1232
    [Google Scholar]
  12. Guyer M.S. 1978; The γδ sequence of F is an insertion sequence.. Journal of Molecular Biology 126:347–365
    [Google Scholar]
  13. Hall F.A. 1971; Bacteriocine typing of Klebsiella spp.. Journal of Clinical Pathology 24:712–716
    [Google Scholar]
  14. Heddell G.W., Mitchell A.B. 1978; Evaluation and application of an improved bacteriocin typing method for Klebsiella aerogenes.. Journal of Clinical Pathology 31:16–21
    [Google Scholar]
  15. Herschman H.R., Helinski D.R. 1967; Comparative study of the events associated with colicin induction.. Journal of Bacteriology 94:691–699
    [Google Scholar]
  16. Israil A.M. 1981; The significance of bacteriocin typing of Klebsiella strains.. Zentralblatt für Bakteriologie und Hygiene(Abteilung I, Originate A) 250:63–71
    [Google Scholar]
  17. James R., Schneider J., Cooper P.C. 1987; Characterization of three group A klebicinplasmids : localization of their E colicin immunity genes.. Journal of General Microbiology 133:2253–2262
    [Google Scholar]
  18. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature; London: 227680–685
    [Google Scholar]
  19. Males B.M., Stocker B.A.D. 1982; ColicinsE4, E5, E6 and A and properties of btuB+ colicinogenic transconjugants.. Journal of General Microbiology 128:95–106
    [Google Scholar]
  20. Maniatis T., Fritsch E.F., Sambrook J. 1982 Molecular Cloning: a laboratory Manual. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  21. Miller J. 1972 Experiments in Molecular Genetics. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  22. Mock M., Miyada C.G., Gunsalus R.P. 1983; Nucleotide sequence for the catalytic domain of colicin E3 and its immunity protein.Evidence for a third gene overlapping colicin.. Nucleic Acids Research 11:3547–3557
    [Google Scholar]
  23. Ørskov I., Ørskov F. 1984; Serotyping of Klebsiella.. Methods in Microbiology 14:143–164
    [Google Scholar]
  24. Ozeki H., Stocker B.A.D., Smith S.M. 1962; Transmission of colicinogeny between strains of Salmonella typhimurium grown together.. Journal of General Microbiology 28:671–687
    [Google Scholar]
  25. Pugsley A.P., Oudega B. 1987; Methods for studying colicins and their plasmids.. In Plasmids: A Practical Approach. Hardy K. G. Edited by Oxford: IRL Press;
    [Google Scholar]
  26. Pugsley A.P., Schwartz M. 1983; A genetic approach to the study of mitomycin induced lysis of Escherichia coli K12 strains which produce colicin E2.. Molecular and General Genetics 190:366–372
    [Google Scholar]
  27. Shafferman A., Flashner Y., Cohen S. 1979; ColEl DNA sequences interacting in cis, essential for mitomycin C induced lethality.. Molecular and General Genetics 176:139–146
    [Google Scholar]
  28. Slopek S., Maresz-Babczyszyn J. 1967; A working scheme for typing Klebsiella bacilli by means of pneumocins.. Archivum immunologiae et therapie experimentalis 15:525–529
    [Google Scholar]
  29. Suit J.L., Fan M.-L.J., Sabik J.F., Labarre R., Luria S.E. 1983; Alternative forms of lethality in mitomycin C-induced bacteria carrying ColE1 plasmids.. Proceedings of the National Academy of Sciences of the United States of America 80:579–583
    [Google Scholar]
  30. Vieira J., Messing J. 1982; The pUC plasmids, an M13mp7 derived system for insertion mutagenesis and sequencing with synthetic universal primers.. Gene 19:259–268
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-134-9-2525
Loading
/content/journal/micro/10.1099/00221287-134-9-2525
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