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. 1996 Mar;62(3):791–797. doi: 10.1128/aem.62.3.791-797.1996

Analysis of the enterohemorrhagic Escherichia coli O157 DNA region containing lambdoid phage gene p and Shiga-like toxin structural genes.

M Datz 1, C Janetzki-Mittmann 1, S Franke 1, F Gunzer 1, H Schmidt 1, H Karch 1
PMCID: PMC167845  PMID: 8975608

Abstract

In this study, we determined the nucleotide sequence of the p gene contained within a 5-kb EcoRI restriction fragment cloned from Shiga-like toxin II (SLT-II)-converting phage 933W of Escherichia coli O157:H7 strain EDL933. The p gene was 702 bp long and had 95.3% sequence similarity to the p gene of phage lambda. Multiple hybridization patterns were obtained when genomic DNA fragments were hybridized with both p and slt-I, slt-II, or slt-IIc sequences. All O157 isolates also possessed an analog of lambda gene p which was not linked with either slt-I or slt-II. Restriction fragment length polymorphism comparisons of clinical O157 isolates and derivates undergoing genotype turnover during infection were made, and loss of large DNA fragments that hybridized with slt-II and p sequences was observed. To further analyze the DNA region containing the p and slt genes, we amplified fragments by using a PCR with one primer complementary to p and the other complementary to either the slt-I or the slt-II gene. PCR analysis with enterohemorrhagic E. coli O157 and non-O157 strains yielded PCR products that varied in size between 5.1 and 7.8 kb. These results suggest that even within O157 isolates, the genomes of SLT-converting phages differ. The methods described here may assist in further investigation of SLT-encoding phages and their role in the epidemiology of infection with enterohemorrhagic E. coli.

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Selected References

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  1. Ahmed R., Bopp C., Borczyk A., Kasatiya S. Phage-typing scheme for Escherichia coli O157:H7. J Infect Dis. 1987 Apr;155(4):806–809. doi: 10.1093/infdis/155.4.806. [DOI] [PubMed] [Google Scholar]
  2. Appleyard R K. Segregation of New Lysogenic Types during Growth of a Doubly Lysogenic Strain Derived from Escherichia Coli K12. Genetics. 1954 Jul;39(4):440–452. doi: 10.1093/genetics/39.4.440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Böhm H., Karch H. DNA fingerprinting of Escherichia coli O157:H7 strains by pulsed-field gel electrophoresis. J Clin Microbiol. 1992 Aug;30(8):2169–2172. doi: 10.1128/jcm.30.8.2169-2172.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Griffin P. M., Tauxe R. V. The epidemiology of infections caused by Escherichia coli O157:H7, other enterohemorrhagic E. coli, and the associated hemolytic uremic syndrome. Epidemiol Rev. 1991;13:60–98. doi: 10.1093/oxfordjournals.epirev.a036079. [DOI] [PubMed] [Google Scholar]
  5. Hii J. H., Gyles C., Morooka T., Karmali M. A., Clarke R., De Grandis S., Brunton J. L. 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. J Clin Microbiol. 1991 Dec;29(12):2704–2709. doi: 10.1128/jcm.29.12.2704-2709.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Huang A., Friesen J., Brunton J. L. Characterization of a bacteriophage that carries the genes for production of Shiga-like toxin 1 in Escherichia coli. J Bacteriol. 1987 Sep;169(9):4308–4312. doi: 10.1128/jb.169.9.4308-4312.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Karch H., Böhm H., Schmidt H., Gunzer F., Aleksic S., Heesemann J. Clonal structure and pathogenicity of Shiga-like toxin-producing, sorbitol-fermenting Escherichia coli O157:H-. J Clin Microbiol. 1993 May;31(5):1200–1205. doi: 10.1128/jcm.31.5.1200-1205.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Karch H., Rüssmann H., Schmidt H., Schwarzkopf A., Heesemann J. Long-term shedding and clonal turnover of enterohemorrhagic Escherichia coli O157 in diarrheal diseases. J Clin Microbiol. 1995 Jun;33(6):1602–1605. doi: 10.1128/jcm.33.6.1602-1605.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Newland J. W., Neill R. J. DNA probes for Shiga-like toxins I and II and for toxin-converting bacteriophages. J Clin Microbiol. 1988 Jul;26(7):1292–1297. doi: 10.1128/jcm.26.7.1292-1297.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. O'Brien A. D., Newland J. W., Miller S. F., Holmes R. K., Smith H. W., Formal S. B. Shiga-like toxin-converting phages from Escherichia coli strains that cause hemorrhagic colitis or infantile diarrhea. Science. 1984 Nov 9;226(4675):694–696. doi: 10.1126/science.6387911. [DOI] [PubMed] [Google Scholar]
  11. Ostroff S. M., Tarr P. I., Neill M. A., Lewis J. H., Hargrett-Bean N., Kobayashi J. M. Toxin genotypes and plasmid profiles as determinants of systemic sequelae in Escherichia coli O157:H7 infections. J Infect Dis. 1989 Dec;160(6):994–998. doi: 10.1093/infdis/160.6.994. [DOI] [PubMed] [Google Scholar]
  12. Paros M., Tarr P. I., Kim H., Besser T. E., Hancock D. D. A comparison of human and bovine Escherichia coli O157:H7 isolates by toxin genotype, plasmid profile, and bacteriophage lambda-restriction fragment length polymorphism profile. J Infect Dis. 1993 Nov;168(5):1300–1303. doi: 10.1093/infdis/168.5.1300. [DOI] [PubMed] [Google Scholar]
  13. Pridmore R. D. New and versatile cloning vectors with kanamycin-resistance marker. Gene. 1987;56(2-3):309–312. doi: 10.1016/0378-1119(87)90149-1. [DOI] [PubMed] [Google Scholar]
  14. Rietra P. J., Willshaw G. A., Smith H. R., Field A. M., Scotland S. M., Rowe B. Comparison of Vero-cytotoxin-encoding phages from Escherichia coli of human and bovine origin. J Gen Microbiol. 1989 Aug;135(8):2307–2318. doi: 10.1099/00221287-135-8-2307. [DOI] [PubMed] [Google Scholar]
  15. Rüssmann H., Kothe E., Schmidt H., Franke S., Harmsen D., Caprioli A., Karch H. Genotyping of Shiga-like toxin genes in non-O157 Escherichia coli strains associated with haemolytic uraemic syndrome. J Med Microbiol. 1995 Jun;42(6):404–410. doi: 10.1099/00222615-42-6-404. [DOI] [PubMed] [Google Scholar]
  16. Rüssmann H., Schmidt H., Caprioli A., Karch H. Highly conserved B-subunit genes of Shiga-like toxin II variants found in Escherichia coli O157 strains. FEMS Microbiol Lett. 1994 May 15;118(3):335–340. doi: 10.1111/j.1574-6968.1994.tb06849.x. [DOI] [PubMed] [Google Scholar]
  17. Rüssmann H., Schmidt H., Heesemann J., Caprioli A., Karch H. Variants of Shiga-like toxin II constitute a major toxin component in Escherichia coli O157 strains from patients with haemolytic uraemic syndrome. J Med Microbiol. 1994 May;40(5):338–343. doi: 10.1099/00222615-40-5-338. [DOI] [PubMed] [Google Scholar]
  18. Samadpour M., Grimm L. M., Desai B., Alfi D., Ongerth J. E., Tarr P. I. Molecular epidemiology of Escherichia coli O157:H7 strains by bacteriophage lambda restriction fragment length polymorphism analysis: application to a multistate foodborne outbreak and a day-care center cluster. J Clin Microbiol. 1993 Dec;31(12):3179–3183. doi: 10.1128/jcm.31.12.3179-3183.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sanger F., Coulson A. R., Hong G. F., Hill D. F., Petersen G. B. Nucleotide sequence of bacteriophage lambda DNA. J Mol Biol. 1982 Dec 25;162(4):729–773. doi: 10.1016/0022-2836(82)90546-0. [DOI] [PubMed] [Google Scholar]
  20. Smith H. W., Green P., Parsell Z. Vero cell toxins in Escherichia coli and related bacteria: transfer by phage and conjugation and toxic action in laboratory animals, chickens and pigs. J Gen Microbiol. 1983 Oct;129(10):3121–3137. doi: 10.1099/00221287-129-10-3121. [DOI] [PubMed] [Google Scholar]
  21. Strockbine N. A., Marques L. R., Newland J. W., Smith H. W., Holmes R. K., O'Brien A. D. Two toxin-converting phages from Escherichia coli O157:H7 strain 933 encode antigenically distinct toxins with similar biologic activities. Infect Immun. 1986 Jul;53(1):135–140. doi: 10.1128/iai.53.1.135-140.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Thomas A., Smith H. R., Rowe B. Use of digoxigenin-labelled oligonucleotide DNA probes for VT2 and VT2 human variant genes to differentiate Vero cytotoxin-producing Escherichia coli strains of serogroup O157. J Clin Microbiol. 1993 Jul;31(7):1700–1703. doi: 10.1128/jcm.31.7.1700-1703.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wang G., Whittam T. S., Berg C. M., Berg D. E. RAPD (arbitrary primer) PCR is more sensitive than multilocus enzyme electrophoresis for distinguishing related bacterial strains. Nucleic Acids Res. 1993 Dec 25;21(25):5930–5933. doi: 10.1093/nar/21.25.5930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Whittam T. S., Wachsmuth I. K., Wilson R. A. Genetic evidence of clonal descent of Escherichia coli O157:H7 associated with hemorrhagic colitis and hemolytic uremic syndrome. J Infect Dis. 1988 Jun;157(6):1124–1133. doi: 10.1093/infdis/157.6.1124. [DOI] [PubMed] [Google Scholar]
  25. Willshaw G. A., Smith H. R., Scotland S. M., Field A. M., Rowe B. Heterogeneity of Escherichia coli phages encoding Vero cytotoxins: comparison of cloned sequences determining VT1 and VT2 and development of specific gene probes. J Gen Microbiol. 1987 May;133(5):1309–1317. doi: 10.1099/00221287-133-5-1309. [DOI] [PubMed] [Google Scholar]

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