DOI QR코드

DOI QR Code

Cloning and Characterization of the Tetracycline Resistant Gene, tetB, from Vibrio parahaemolyticus

  • Kang, Min-Seung (Department of Food Science and Technology, Pukyong National University) ;
  • Park, Kun-Ba-Wui (South Sea Fisheries Research Institute, National Fisheries Research & Development Institute) ;
  • Hwang, Hye-Jin (Department of Food Science and Technology, Pukyong National University) ;
  • Bae, Hyang-Nam (Department of Food Science and Technology, Pukyong National University) ;
  • Lim, Keun-Sik (Department of Food Science and Technology, Pukyong National University) ;
  • Eom, Sung-Hwan (Department of Food Science and Technology, Pukyong National University) ;
  • Kim, Young-Mog (Department of Food Science and Technology, Pukyong National University)
  • Published : 2009.03.31

Abstract

A tetracycline resistant Vibrio parahaemolyticus, capable of growing on TCBS medium containing tetracycline, was isolated from cultivated fishes. A gene responsible for the tetracycline resistance was cloned from chromosomal DNA of the V. parahaemolyticus strain using Escherichia coli KAM3, which lacks major multi-drug efflux pumps (${\Delta}acrB$) as host cells. The nucleotide sequence and homology analysis revealed an open reading frame (ORF) for tetracycline resistance protein (TetB). In order to characterize the antibiotic resistance of TetB originated from the V. parahaemolyticus strain, the gene was sub cloned into plasmid pSTV28. The resulting plasmid was designated as pSTVTetB and transformated into E. coli KAM3. E. coli KAM3 cells harboring the recombinant plasmid pSTVTetB are able to grow on plates containing tetracycline and oxytetracycline but not doxycycline, indicating that the tetB gene confers the tetracycline- and oxytetracycline-resistance to the host cell.

Keywords

References

  1. Andersen, S.R. and R.A. Sandra. 1994. Distribution of tetracycline resistance determinants among gramnegative bacteria isolated from polluted and unpol1uted marine sediments. Appl. Environ. Microbiol., 60, 908-912
  2. Depaola, A. and M.C. Roberts. 1995. Class D and E tetracycline resistance determinants in gram-negative bacteria from catfish ponds. Mol. Cell Probes., 9, 311-313 https://doi.org/10.1016/S0890-8508(95)91572-9
  3. Depaola, A., W.E. Hill and F.M. Harrell. 1993. Oligonucleotide probe determination of tetracycline resistant bacteria isolated from catfish ponds. Mol. Cell Probes., 7, 345-348 https://doi.org/10.1006/mcpr.1993.1051
  4. Furushita, M., T. Shiba, T. Maeda, M. Yahata, A. Kaneoka, Y. Takahashi, K. Torii, T. Hasegawa and M. Ohta. 2003. Similarity of tetracycline resistance genes isolated from fish farm bacteria to those from clinical isolates. Appl. Environ. Microbiol., 69, 5336-5342 https://doi.org/10.1128/AEM.69.9.5336-5342.2003
  5. Jeanette,W., P. Teo, M.C. Tan Theresa and L.P. Chit. 2002. Genεtic determinants of tetracycline resistance in Vibrio harveyi. Antimicro. Agents Chemother.,46, 1038-1045 https://doi.org/10.1128/AAC.46.4.1038-1045.2002
  6. Jun, L.J., J.B. Jeong, J.H. Kim, C.H. Lee, E.H. Kim and H.D. Jeong. 2007. Distribution of tet genes in the tetracycline resistant microorganisms isolated from marin-aquatic environments of Korea. l. Fish Pathol.,20, 169-178
  7. Kim, Y.B. 2001. Virulence factors and genotyping of Vibrio parahaemolyticus in sea water and clinical isolates. J. Bacteriol. Virol., 31, 229-238
  8. Kim, Y.H., L.J. Jun, S.H. Park, S.H. Yoon and J.K. Chung. 2007. Prevalence of tet(B) and tet(M) genes among tetracyc1ine-resistant Vibrio spp. in the aquatic environments of Korea. Dis. Aquat. Org., 75, 209-216 https://doi.org/10.3354/dao075209
  9. National Committee for Clinical Laboratory Standards. 2002. Performance standards for antimicrobial disk susceptibility test. National Committee for Clinical Laboratory Standards. Wayne, PA
  10. Nonaka, L. and S. Suzuki. 2002. New $MgL^+$-dependent oxytetracycline resistance determinant tet34 in Vibrio isolates from marine fish intestinal contents. Antimicrob. Agents Chemother., 476, 1550-1552 https://doi.org/10.1128/AAC.46.5.1550-1552
  11. Pratt, A. and V. Korolik. 2005. Tetracyc1ine resistance of Australian Campylobacter jejuni and Campylobacter coli isolstes. J. Antimicrob. Agents Chemother., 55, 452-460 https://doi.org/10.1093/jac/dki040
  12. Roberts, M.C. 2005. Tetracycline resistance determinants: mechanisms of action, regulation of expression, genetic mobility, and distribution. FEMS Microbiol. Rev., 19, 1-24
  13. Sanger, F., S. Nicklen and A.R. Coulson. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA., 74, 5463-5467 https://doi.org/10.1073/pnas.74.12.5463
  14. Schmidt, A. S., M.S. Bruun, I. Dalsgaard and J.L. Larsen 2001. Incidence, distribution and spread of tetracycline resistance determinants and integron -associated antibiotic resistance genes among motile aeromonads from a fish farming environment. Appl. Environ. Microbiol., 67, 5675-5682 https://doi.org/10.1128/AEM.67.12.5675-5682.2001
  15. Teo, J.W.P., T.M.C. Tan and C.L. Poh. 2002. Genetic determinants of tetracycline resistance in Vibrio harveyi. antimicrob. Agents Chemother., 46, 1038-1045 https://doi.org/10.1128/AAC.46.4.1038-1045.2002