Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1985 Nov;50(2):420–424. doi: 10.1128/iai.50.2.420-424.1985

Plasmid-mediated virulence in Salmonella dublin demonstrated by use of a Tn5-oriT construct.

G K Chikami, J Fierer, D G Guiney
PMCID: PMC261968  PMID: 2997039

Abstract

Salmonella dublin, a serotype which causes invasive disease in cattle and humans, carries a characteristic 80-kilobase plasmid (pSDL2). We were able to cure the plasmid from a strain of S. dublin. The cured strain was avirulent for mice by either the oral or intraperitoneal route of infection. A derivative of Tn5 which contains the transfer origin of the broad-host-range plasmid RK2 (Tn5-oriT) was transposed onto pSDL2, allowing mobilization of the plasmid by an RK2 helper plasmid. Reintroduction of the pSDL2 derivative plasmid into the cured strain restored virulence, demonstrating that the plasmid is necessary for virulence. These studies also demonstrate the usefulness of the Tn5-oriT construct for genetic manipulations.

Full text

PDF
420

Images in this article

Selected References

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

  1. Bochner B. R., Huang H. C., Schieven G. L., Ames B. N. Positive selection for loss of tetracycline resistance. J Bacteriol. 1980 Aug;143(2):926–933. doi: 10.1128/jb.143.2.926-933.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Bouanchaud D. H., Scavizzi M. R., Chabbert Y. A. Elimination by ethidium bromide of antibiotic resistance in enterobacteria and staphylococci. J Gen Microbiol. 1968 Dec;54(3):417–425. doi: 10.1099/00221287-54-3-417. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Currier T. C., Nester E. W. Isolation of covalently closed circular DNA of high molecular weight from bacteria. Anal Biochem. 1976 Dec;76(2):431–441. doi: 10.1016/0003-2697(76)90338-9. [DOI] [PubMed] [Google Scholar]
  6. Elwell L. P., Shipley P. L. Plasmid-mediated factors associated with virulence of bacteria to animals. Annu Rev Microbiol. 1980;34:465–496. doi: 10.1146/annurev.mi.34.100180.002341. [DOI] [PubMed] [Google Scholar]
  7. Fierer J., Fleming W. Distinctive biochemical features of Salmonella dublin isolated in California. J Clin Microbiol. 1983 Mar;17(3):552–554. doi: 10.1128/jcm.17.3.552-554.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fierer J. Invasive Salmonella dublin infections associated with drinking raw milk. West J Med. 1983 May;138(5):665–669. [PMC free article] [PubMed] [Google Scholar]
  9. Figurski D. H., Helinski D. R. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1648–1652. doi: 10.1073/pnas.76.4.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Guiney D. G., Hasegawa P., Davis C. E. Plasmid transfer from Escherichia coli to Bacteroides fragilis: differential expression of antibiotic resistance phenotypes. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7203–7206. doi: 10.1073/pnas.81.22.7203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Guiney D. G., Yakobson E. Location and nucleotide sequence of the transfer origin of the broad host range plasmid RK2. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3595–3598. doi: 10.1073/pnas.80.12.3595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hall G. A., Jones P. W., Aitken M. M. The pathogenesis of experimental intra-ruminal infections of cows with Salmonella dublin. J Comp Pathol. 1978 Jul;88(3):409–417. doi: 10.1016/0021-9975(78)90045-2. [DOI] [PubMed] [Google Scholar]
  13. Harris J. R., Wachsmuth I. K., Davis B. R., Cohen M. L. High-molecular-weight plasmid correlates with Escherichia coli enteroinvasiveness. Infect Immun. 1982 Sep;37(3):1295–1298. doi: 10.1128/iai.37.3.1295-1298.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Helmuth R., Stephan R., Bunge C., Hoog B., Steinbeck A., Bulling E. Epidemiology of virulence-associated plasmids and outer membrane protein patterns within seven common Salmonella serotypes. Infect Immun. 1985 Apr;48(1):175–182. doi: 10.1128/iai.48.1.175-182.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hershfield V., Boyer H. W., Yanofsky C., Lovett M. A., Helinski D. R. Plasmid ColEl as a molecular vehicle for cloning and amplification of DNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3455–3459. doi: 10.1073/pnas.71.9.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jones G. W., Rabert D. K., Svinarich D. M., Whitfield H. J. Association of adhesive, invasive, and virulent phenotypes of Salmonella typhimurium with autonomous 60-megadalton plasmids. Infect Immun. 1982 Nov;38(2):476–486. doi: 10.1128/iai.38.2.476-486.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kopecko D. J., Washington O., Formal S. B. Genetic and physical evidence for plasmid control of Shigella sonnei form I cell surface antigen. Infect Immun. 1980 Jul;29(1):207–214. doi: 10.1128/iai.29.1.207-214.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McHugh G. L., Swartz M. N. Elimination of plasmids from several bacterial species by novobiocin. Antimicrob Agents Chemother. 1977 Sep;12(3):423–426. doi: 10.1128/aac.12.3.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nakamura M., Sato S., Ohya T., Suzuki S., Ikeda S. Possible relationship of a 36-megadalton Salmonella enteritidis plasmid to virulence in mice. Infect Immun. 1985 Mar;47(3):831–833. doi: 10.1128/iai.47.3.831-833.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Portnoy D. A., Wolf-Watz H., Bolin I., Beeder A. B., Falkow S. Characterization of common virulence plasmids in Yersinia species and their role in the expression of outer membrane proteins. Infect Immun. 1984 Jan;43(1):108–114. doi: 10.1128/iai.43.1.108-114.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sansonetti P. J., Kopecko D. J., Formal S. B. Involvement of a plasmid in the invasive ability of Shigella flexneri. Infect Immun. 1982 Mar;35(3):852–860. doi: 10.1128/iai.35.3.852-860.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sigeti J. S., Guiney D. G., Jr, Davis C. E. Mechanism of action of metronidazole on Bacteroides fragilis. J Infect Dis. 1983 Dec;148(6):1083–1089. doi: 10.1093/infdis/148.6.1083. [DOI] [PubMed] [Google Scholar]
  23. Terakado N., Sekizaki T., Hashimoto K., Naitoh S. Correlation between the presence of a fifty-megadalton plasmid in Salmonella dublin and virulence for mice. Infect Immun. 1983 Jul;41(1):443–444. doi: 10.1128/iai.41.1.443-444.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Thomas C. M. Complementation analysis of replication and maintenance functions of broad host range plasmids RK2 and RP1. Plasmid. 1981 May;5(3):277–291. doi: 10.1016/0147-619x(81)90005-6. [DOI] [PubMed] [Google Scholar]
  25. Watanabe H., Timmis K. N. A small plasmid in Shigella dysenteriae 1 specifies one or more functions essential for O antigen production and bacterial virulence. Infect Immun. 1984 Jan;43(1):391–396. doi: 10.1128/iai.43.1.391-396.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Werner S. B., Humphrey G. L., Kamei I. Association between raw milk and human Salmonella dublin infection. Br Med J. 1979 Jul 28;2(6184):238–241. doi: 10.1136/bmj.2.6184.238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yakobson E. A., Guiney D. G., Jr Conjugal transfer of bacterial chromosomes mediated by the RK2 plasmid transfer origin cloned into transposon Tn5. J Bacteriol. 1984 Oct;160(1):451–453. doi: 10.1128/jb.160.1.451-453.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Zink D. L., Feeley J. C., Wells J. G., Vanderzant C., Vickery J. C., Roof W. D., O'Donovan G. A. Plasmid-mediated tissue invasiveness in Yersinia enterocolitica. Nature. 1980 Jan 10;283(5743):224–226. doi: 10.1038/283224a0. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES