A 3.3-kb plasmid of enterohemorrhagic Escherichia coli O157:H7 is closely related to the core region of the Salmonella typhimurium antibiotic resistance plasmid NTP16.

A restriction enzyme map was constructed of a 3.3-kb plasmid derived from enterohemorrhagic Escherichia coli O157:H7 strain 4821 using the enzymes SmaI, HpaI, FokI, HaeIII, StyI, RsaI, Bg/II, ClaI, and EcoRV. The molecular size of p4821 was 3307 bp, determined by nucleotide sequencing. Homology searches in the EMBL database library revealed that the nucleotide sequence of P4821 is similar (> 98%) to the core region of the antibiotic resistance plasmid NTP16 of Salmonella typhimurium strains. Nucleotide sequence analysis showed that p4821 contains all the information necessary for its replication, stability, and mobilization. However, the lack of tra genes indicates that the plasmid is nonconjugative. A possible transposon insertion site was found in p4821 but two such sites were found at the boundaries between the core region and the antibiotic resistance encoding transposons in plasmid NTP16. Using a hybridization assay, we determined that of 50 E. coli O157:H7 strains isolated in 1996 in Würzburg, Germany from patients with diarrhea and hemolytic-uremic syndrome, 4 strains (8%) contained plasmid p4821-specific sequences.

[1]  H. Karch,et al.  Structure and Function of Plasmid pColD157 of Enterohemorrhagic Escherichia coli O157 and Its Distribution among Strains from Patients with Diarrhea and Hemolytic-Uremic Syndrome , 1998, Journal of Clinical Microbiology.

[2]  H. Karch,et al.  EspP, a novel extracellular serine protease of enterohaemorrhagic Escherichia coli O157:H7 cleaves human coagulation factor V , 1997, Molecular microbiology.

[3]  H. Karch,et al.  A gene cluster closely related to type II secretion pathway operons of gram-negative bacteria is located on the large plasmid of enterohemorrhagic Escherichia coli O157 strains. , 1997, FEMS microbiology letters.

[4]  H. Karch,et al.  KatP, a novel catalase-peroxidase encoded by the large plasmid of enterohaemorrhagic Escherichia coli O157:H7. , 1996, Microbiology.

[5]  H. Karch,et al.  Analysis of the EHEC hly operon and its location in the physical map of the large plasmid of enterohaemorrhagic Escherichia coli O157:h7. , 1996, Microbiology.

[6]  J. Meng,et al.  Molecular characterisation of Escherichia coli O157:H7 isolates by pulsed-field gel electrophoresis and plasmid DNA analysis. , 1995, Journal of medical microbiology.

[7]  L. Beutin,et al.  Molecular analysis of the plasmid-encoded hemolysin of Escherichia coli O157:H7 strain EDL 933 , 1995, Infection and immunity.

[8]  T. McDaniel,et al.  A genetic locus of enterocyte effacement conserved among diverse enterobacterial pathogens. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[9]  C. Hutchison,et al.  Insertion site specificity of the transposon Tn3. , 1995, Nucleic acids research.

[10]  T. Besser,et al.  A comparison of human and bovine Escherichia coli O157:H7 isolates by toxin genotype, plasmid profile, and bacteriophage lambda-restriction fragment length polymorphism profile. , 1993, The Journal of infectious diseases.

[11]  H. Karch,et al.  DNA fingerprinting of Escherichia coli O157:H7 strains by pulsed-field gel electrophoresis , 1992, Journal of clinical microbiology.

[12]  P. Cannon,et al.  Complete nucleotide sequence and gene organization of plasmid NTP16. , 1992, Plasmid.

[13]  M. P. Jackson,et al.  Shiga toxin: biochemistry, genetics, mode of action, and role in pathogenesis. , 1992, Current topics in microbiology and immunology.

[14]  H. Lior,et al.  Colicinogeny of O157:H7 enterohemorrhagic Escherichia coli and the shielding of colicin and phage receptors by their O-antigenic side chains. , 1991, Canadian journal of microbiology.

[15]  R. Tauxe,et al.  The epidemiology of infections caused by Escherichia coli O157:H7, other enterohemorrhagic E. coli, and the associated hemolytic uremic syndrome. , 1991, Epidemiologic reviews.

[16]  J. Tomizawa.  Control of ColE1 plasmid replication. Interaction of Rom protein with an unstable complex formed by RNA I and RNA II. , 1990, Journal of molecular biology.

[17]  J. Tomizawa.  Control of ColE1 plasmid replication. Intermediates in the binding of RNA I and RNA II. , 1990, Journal of molecular biology.

[18]  S. Ostroff,et al.  Toxin genotypes and plasmid profiles as determinants of systemic sequelae in Escherichia coli O157:H7 infections. , 1989, The Journal of infectious diseases.

[19]  J. Wells,et al.  Illnesses associated with Escherichia coli O157:H7 infections. A broad clinical spectrum. , 1988, Annals of internal medicine.

[20]  S. Ratnam,et al.  Characterization of Escherichia coli serotype O157:H7 , 1988, Journal of clinical microbiology.

[21]  D. Sherratt,et al.  Resolution of ColE1 dimers requires a DNA sequence implicated in the three‐dimensional organization of the cer site. , 1988, The EMBO journal.

[22]  M. Levine,et al.  A DNA probe to identify enterohemorrhagic Escherichia coli of O157:H7 and other serotypes that cause hemorrhagic colitis and hemolytic uremic syndrome. , 1987, The Journal of infectious diseases.

[23]  P. Strike,et al.  Characterization of the drug resistance plasmid NTP16. , 1987, Plasmid.

[24]  J. Lebowitz,et al.  Nucleotide sequence and gene organization of ColE1 DNA. , 1985, The Journal of biological chemistry.

[25]  D. Sherratt,et al.  Multimerization of high copy number plasmids causes instability: Cole 1 encodes a determinant essential for plasmid monomerization and stability , 1984, Cell.

[26]  Regulation of plasmid replication. , 1984, Microbiological reviews.

[27]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[28]  A. Clark,et al.  Conjugal transmission of plasmids. , 1979, Annual review of genetics.

[29]  S. Cohen,et al.  Studies of the specificity and control of transposition of the Tn3 element. , 1979, Cold Spring Harbor symposia on quantitative biology.