Skip to main content
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1992 Mar;36(3):611–615. doi: 10.1128/aac.36.3.611

Identification and cloning of a tetracycline resistance gene from the fish pathogen Vibrio salmonicida.

H Sørum 1, M C Roberts 1, J H Crosa 1
PMCID: PMC190565  PMID: 1622171

Abstract

Vibrio salmonicida is the causative agent of cold-water vibriosis in farmed Atlantic salmon. We cloned a 6.9-kb PstI fragment from the 170-MDa plasmid (pRVS1) containing a tetracycline resistance determinant. A subcloned 1.96-kb HindIII fragment was found to mediate the tetracycline resistance. The 2.5-kb ClaI-PvuI fragment carrying the Tet E determinant from Escherichia coli hybridized under stringent conditions with the cloned 1.96-kb HindIII fragment from V. salmonicida. The 1.96-kb HindIII fragment codes for a protein of 26.5 kDa which represents a candidate for the structural TetA protein of the class E determinant. Deletion of a 0.28-kb BalI fragment from the middle of the HindIII fragment resulted in the loss of tetracycline resistance. We were able to show that when E. coli carries the cloned 6.9-kb PstI fragment, expression of tetracycline resistance is regulated by the concentration of tetracycline in the medium. In contrast, tetracycline resistance was constitutively expressed in the E. coli isolate carrying the 1.96-kb HindIII fragment. The tetracycline resistance gene isolated from the 170-MDa R plasmid of the marine fish pathogen V. salmonicida was characterized and shown to be a class E determinant.

Full text

PDF
611

Images in this article

Selected References

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

  1. Aoki T. Drug-resistant plasmids from fish pathogens. Microbiol Sci. 1988 Jul;5(7):219–223. [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. 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]
  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. 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]
  6. DePaola A., Flynn P. A., McPhearson R. M., Levy S. B. Phenotypic and genotypic characterization of tetracycline- and oxytetracycline-resistant Aeromonas hydrophila from cultured channel catfish (Ictalurus punctatus) and their environments. Appl Environ Microbiol. 1988 Jul;54(7):1861–1863. doi: 10.1128/aem.54.7.1861-1863.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Levy S. B., Buu-Hoi A., Marshall B. Transposon Tn10-like tetracycline resistance determinants in Haemophilus parainfluenzae. J Bacteriol. 1984 Oct;160(1):87–94. doi: 10.1128/jb.160.1.87-94.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Levy S. B., McMurry L. M., Burdett V., Courvalin P., Hillen W., Roberts M. C., Taylor D. E. Nomenclature for tetracycline resistance determinants. Antimicrob Agents Chemother. 1989 Aug;33(8):1373–1374. doi: 10.1128/aac.33.8.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Marshall B., Morrissey S., Flynn P., Levy S. B. A new tetracycline-resistance determinant, class E, isolated from Enterobacteriaceae. Gene. 1986;50(1-3):111–117. doi: 10.1016/0378-1119(86)90315-x. [DOI] [PubMed] [Google Scholar]
  11. Marshall B., Roberts M., Smith A., Levy S. B. Homogeneity of transferable tetracycline-resistance determinants in Haemophilus species. J Infect Dis. 1984 Jun;149(6):1028–1029. doi: 10.1093/infdis/149.6.1028. [DOI] [PubMed] [Google Scholar]
  12. Marshall B., Tachibana C., Levy S. B. Frequency of tetracycline resistance determinant classes among lactose-fermenting coliforms. Antimicrob Agents Chemother. 1983 Dec;24(6):835–840. doi: 10.1128/aac.24.6.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mendez B., Tachibana C., Levy S. B. Heterogeneity of tetracycline resistance determinants. Plasmid. 1980 Mar;3(2):99–108. doi: 10.1016/0147-619x(80)90101-8. [DOI] [PubMed] [Google Scholar]
  14. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  15. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  16. Søorum H., Poppe T. T., Olsvik O. Plasmids in Vibrio salmonicida isolated from salmonids with hemorrhagic syndrome (Hitra disease). J Clin Microbiol. 1988 Sep;26(9):1679–1683. doi: 10.1128/jcm.26.9.1679-1683.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sørum H., Hvaal A. B., Heum M., Daae F. L., Wiik R. Plasmid profiling of Vibrio salmonicida for epidemiological studies of cold-water vibriosis in Atlantic salmon (Salmo salar) and cod (Gadus morhua). Appl Environ Microbiol. 1990 Apr;56(4):1033–1037. doi: 10.1128/aem.56.4.1033-1037.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tovar K., Ernst A., Hillen W. Identification and nucleotide sequence of the class E tet regulatory elements and operator and inducer binding of the encoded purified Tet repressor. Mol Gen Genet. 1988 Dec;215(1):76–80. doi: 10.1007/BF00331306. [DOI] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES