Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
권호 표지

Cloning and Nucleotide Sequence Analysis of Verotoxin Gene from Escherichia coli O157 KNIH317 Isolated in Korea

  • Park, Yong-Chjun (School of Life Sciences, Chungbuk National University, Cheongju) ;
  • Shin, Hee-Jung (School of Life Sciences, Chungbuk National University, Cheongju) ;
  • Kim, Young-Chang (School of Life Sciences, Chungbuk National University, Cheongju)
  • Published : 1999.09.01
Download PDF
⟨ Previous Next ⟩

Abstract

Escherichia coli O157 is an important pathogenic organism which causes diarrhea, haemorrhagic colitis, and haemolytic ureamic syndrome (HUS) in human. E. coli O157 KNIH317 was isolated form patients suffering with HUS in Korea. We designed a primer set for cloning shiga-like toxin (slt) gene. The amplified PCR product was used to Southern and colony hybridization as a probe. As a result, we cloned 4.5-kb KpnI fragment containing the slt gene encoding shiga-like toxin from chromosomal DNA of E. coli O157 KNIH317. This recombinant plasmid was named pOVT45. E. coli XL1-Blue harboring pOVT45 showed cytotoxicity in Vero cells. We sequenced the slt gene of this strain. The A-subunit gene of the slt was composed of 960 base pairs with ATG initiation codon and TAA terminationcodon. The B-subunit was composed of 270 base paris with ATG initiation codon and TGA termination codon. Nucleotide sequence comparison of the slt gene exhibited 100%, 98.4%, 93.7%, and 93.7% identity with that of shiga-like toxin type II (sltII) of E. coli bacteriophage 933W, variant slt of E. coli, slt of E. coli, and variant sltII of E. coli, respectively. From these results, it was concluded that the cloned slt gene belongs to SltII family and that the strain used in this study may be a lysogeny of E. coli bcteriphage 933W.

Keywords

References

  1. J. Infect. Dis. v.155 Phage-typing scheme for Escherichia coli O157:H7 Ahmed, R.;C. Bopp;A. Borczyk;S. Kasatiya
  2. Nucl. Acids Res. v.25 Gapped BLAST and PSI-BLAST: a new generation of protein database search programs Altschul, S.F.;T.L. Madden;A.A. Schaffer;J. Zhang;Z. Zhang;W. Miller;D.J. Lipman
  3. J. Clin. Microbiol. v.30 DNA fingerprinting of Escherichia coli O157:H7 strains by pulse-field gel electrophoresis Bohm, H.;H. Karch
  4. Proc. Natl. Acad. Sci. USA v.84 Nucleotide sequence of the shiga-like toxin genes of Escherichia coli Calderwood, S.B.;F. Auclair;A. Donohue-Rolfe;G.T. Keusch;J.J. Mekalanos
  5. J. Bacteriol. v.169 Nucleotide sequence and promoter mapping of the Escherichia coli. Shiga-like toxin operon of bacteriophage H-19B De Grandis, S.;J. Ginsberg;M. Toone;S. Climie;J. Friesen;J. Brunton
  6. J. Clin. Microbiol. v.136 Molecular cloning and nucleotide sequence of another variant of the Escherichia coli Shiga-like toxin Ⅱ family Gannon, V.P.J.;C. Teerling;S.A. Masri;C.L. Gyles
  7. J. Clin. Microbiol. v.29 Development of verotoxin 2- and verotoxin 2 variant (VT2v)-specific oligonucleotide probes on the basis of the nucleotide sequence of the B cistron of VT2v from Escherichia coli E32511 and B2F1 Hii, J.H.;C. Gyles;T. Morooka;M.A. Karmali;R. Clarke;S. De Grandis;J.L. Brunton
  8. FEMS Microbiol. Lett. v.44 Nucleotide sequence analysis and comparison of the structural genes for Shiga-like toxin Ⅰ and Shiga-like toxin Ⅱ encoded by bacteriophages from Escherichia coli 933 Jackson, M.P.;R.J. Neill;A.D. OBrien;R.K. Holmes;J.W. Newland
  9. J. Clin. Microbiol. v.33 Long-term shedding and clonal turnover of enterohemorrhagic Escherichia coli O157 in diarrheal disease Karch, H.;H. R ssmann;H. Schmidt;A. Schwarzkopf;J. Heesemann
  10. Clin. Microbiol. Rev. v.2 Infection by verotoxin-producing Escherichia coli Karmali, M.A.
  11. Gene v.67 The primary structure of the operons coding for Shigella dysenteriae toxin and temperate phage H30 shiga-like toxin Kozlov, Y.V.;A.A. Kabishev;E.V. Lukyanov;A.A. Bayev
  12. J. Clin. Microbiol. v.26 DNA probes for Shiga-like toxins Ⅰ and Ⅱ and for toxin-converting bacteriophages Newland, J.W.;R.J. Neill
  13. J. Infect. Dis. v.160 Toxin genotypes and plasmid profiles as determinants of systemic sequelae in Escherichia coli O157:H7 infections Ostroff, S.M.;P.I. Tarr;M.A. Neill;J.H. Lewis;N. Hargrett Bean;J.M. Kobayashi
  14. Infect. Immun. v.40 Purification and characterization of a Shigella dysenteriae 1-like toxin produced by Escherichia coli O'Brien, A.D.;G.D. La Veck
  15. Microbiol. Rev. v.51 Shiga and shiga-like toxins O'Brien, A.D.;R.K. Holmes
  16. J. Infect. Dis. v.146 Production of Shigella dysenteriae type 1-like cytotoxin by Escherichia coli O'Brien, A.D.;G.D. La Veck;M.R. Thompson;S.B. Formal
  17. Science v.226 Shiga-like toxin-converting phages from Escherichia coli strains that cause hemorrhagic colitis or infantile diarrhea O'Brien, A.D.;J.W. Newland;S.F. Miller;R.K. Holmes;H.W. Smith;S.B. Formal
  18. J. Infect. Dis. v.168 A comparison of human and bovine Escherichia coli O157:H7 isolates by toxin genotype, plasmid profile, and bacteriophage lambda-restriction fragment length polymorphism profile Paros, M.;P.I. Tarr;H. Kim;T.E. Besser;D.D. Hancock
  19. J. Gen. Microbiol. v.135 Comparison of Vero-cytotoxin-encoding phages from Escherichia coli of human and bovine origin Rietra, P.J.G.M.;G.A. Willshaw;H.R. Smith;A.M. Field;S.M. Scotland;B. Rowe
  20. Molecular Cloning: A Laboratory Manual (2nd ed.) Sambrook, J.;E.F. Fritsch;T. Maniatis
  21. Infec Immun. v.59 Two copies of shiga-like toxin Ⅱ-related genes common in enterohemorrhagic Escherichia coli strains are responsible for the antigenic heterogenity of the O157:H- strain E32511 Schmitt, C.K.;M.L. McKee;A.D. OBrien
  22. Lancet ii. Two distinct toxins active Vero cells from Escherichia coli O157 Scotland, S.M.;H.R. Smith;B. Rowe
  23. Infect. Immun. v.53 Two toxin-converting phages from Escherichia coli O157:H7 strain 933 encode antigenically distinct toxins with similar biologic activities Strockbine, N.A.;L.R. Marques;J.W. Newland;H.W. Smith;R.K. Holmes;A.D. OBrien
  24. J. Bacteriol. v.170 Cloning and sequencing of the genes for shiga toxin from Shigella dysenteriae type Ⅰ Strockbine, N.A.;M.P. Jackson;L.M. Sung;R.K. Holmes;A.D. OBrien
  25. Nucleic Acids Res. v.25 The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools Thompson, J.D.;T.J. Gibson;F. Plewniak;F. Jeanmougin;D.G. Higgins
  26. Nucleic Acids Res. v.2157 RAPD (arbitrary primer) PCR is more sensitive than multilocus enzyme electrophoresis for distinguishing related bacterial strains Wang, G.;T.S. Whittam;C.M. Berg;D.E. Berg
  27. J. Infect. Dis. v.157 Genetic evidence of clonal descent of Escherichia coli O157:H7 associated with hemorrhagic colitis and hemolytic uremic syndrome Whittam, T.S.;I.K. Wachsmuth;R.A. Wilson