Characterization of plasmids encoding the adherence factor of enteropathogenic Escherichia coli

Volunteer studies have shown that a 60-megadalton plasmid is required for full virulence of the human enteropathogenic Escherichia coli (EPEC) strain E2348/69 (O127:H6). The plasmid, designated pMAR2, encodes localized adherence to HEp-2 cells in tissue culture via the adhesin known as the EPEC adherence factor (EAF). Using a DNA probe for the EAF, we have previously shown that these genes are specific for EPEC and are usually encoded on plasmids ranging from 55 to 65 megadaltons. In this study, Southern blot analysis and S1 nuclease homology determination reveal a high degree of sequence conservation among these plasmids, despite some variation in restriction maps. Phenotypic characterization of the prototype EAF plasmid pMAR2 reveals that the plasmid belongs to the group IncFII and is negative for alpha-hemolysin, colicin, and aerobactin synthesis, as well as biochemical markers and antibiotic resistance. Regions encoding adherence to HEp-2 cells were localized by Tn801 insertion mutagenesis. Adherence genes were then cloned as two distinct plasmid regions which confer the adherence phenotype only when complementing each other in trans.

[1]  M. Levine,et al.  A plasmid of enterohemorrhagic Escherichia coli O157:H7 is required for expression of a new fimbrial antigen and for adhesion to epithelial cells , 1987, Infection and immunity.

[2]  J. Kaper,et al.  Role of plasmid-encoded adherence factors in adhesion of enteropathogenic Escherichia coli to HEp-2 cells , 1987, Infection and immunity.

[3]  J. Nataro,et al.  Localization of a determinant for HEp-2 adherence by enteropathogenic Escherichia coli , 1986, Infection and immunity.

[4]  I. Scaletsky,et al.  Plasmids coding for drug resistance and localized adherence to HeLa cells in enteropathogenic Escherichia coli O55:H- and O55:H6 , 1986, Infection and immunity.

[5]  D. Galloway,et al.  Cloning of a gene involved in regulation of exotoxin A expression in Pseudomonas aeruginosa , 1986, Infection and immunity.

[6]  P. Blake,et al.  Correlation between adherence to HeLa cells and serogroups, serotypes, and bioserotypes of Escherichia coli , 1985, Infection and immunity.

[7]  M. Levine,et al.  Detection of an adherence factor of enteropathogenic Escherichia coli with a DNA probe. , 1985, The Journal of infectious diseases.

[8]  M. Levine,et al.  Plasmid-mediated factors conferring diffuse and localized adherence of enteropathogenic Escherichia coli , 1985, Infection and immunity.

[9]  P. Williams,et al.  AEROBACTIN-MEDIATED IRON UPTAKE: A VIRULENCE DETERMINANT IN ENTEROPATHOGENIC ESCHERICHIA COLI? , 1985, The Lancet.

[10]  B. Rowe,et al.  Expression of cloned plasmid regions encoding colonization factor antigen I (CFA/I) in Escherichia coli. , 1985, Plasmid.

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

[12]  D. Shlaes,et al.  Variability in DNA sequence of closely related nosocomial gentamicin-resistant plasmids. , 1983, The Journal of infectious diseases.

[13]  M. Levine,et al.  Plasmid‐Mediated Adhesion in Enteropathogenic Escherichia coli , 1983, Journal of pediatric gastroenterology and nutrition.

[14]  M. Levine,et al.  Summary of a workshop on enteropathogenic Escherichia coli. , 1983 .

[15]  A. J. Mcadams,et al.  An ultrastructural study of enteropathogenic Escherichia coli infection in human infants. , 1983, Ultrastructural pathology.

[16]  P. A. Schad,et al.  Characterization of virulence plasmids and plasmid-associated outer membrane proteins in Shigella flexneri, Shigella sonnei, and Escherichia coli , 1983, Infection and immunity.

[17]  P. A. Schad,et al.  Genetic transfer of a mucosal adherence factor (R1) from an enteropathogenic Escherichia coli strain into a Shigella flexneri strain and the phenotypic suppression of this adherence factor. , 1983, The Journal of infectious diseases.

[18]  S. Falkow,et al.  Detection of enterotoxigenic Escherichia coli by DNA colony hybridization. , 1980, The Journal of infectious diseases.

[19]  S. Falkow,et al.  Isolation of a temperature-sensitive derivative of RP1. , 1980, Plasmid.

[20]  M. Ulshen,et al.  Pathogenesis of escherichia coli gastroenteritis in man--another mechanism. , 1980, The New England journal of medicine.

[21]  H. Birnboim,et al.  A rapid alkaline extraction procedure for screening recombinant plasmid DNA. , 1979, Nucleic acids research.

[22]  S. Falkow,et al.  Relationship Between Colicin V Activity and Virulence in Escherichia coli , 1979, Infection and immunity.

[23]  S. Ehrlich,et al.  Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. , 1979, Gene.

[24]  H. Smith,et al.  Further observations on the association of the colicine V plasmid of Escherichia coli with pathogenicity and with survival in the alimentary tract. , 1976, Journal of general microbiology.

[25]  E. Southern.  Detection of specific sequences among DNA fragments separated by gel electrophoresis. , 1975, Journal of molecular biology.

[26]  S. Falkow,et al.  Polynucleotide sequence relationships among Ent plasmids and the relationship between Ent and other plasmids , 1975, Journal of bacteriology.

[27]  S. Falkow,et al.  Use of a Single-Strand Specific Nuclease for Analysis of Bacterial and Plasmid Deoxyribonucleic Acid Homo- and Heteroduplexes , 1973, Journal of bacteriology.

[28]  E. S. Anderson,et al.  Molecular Studies of R Factor Compatibility Groups , 1973, Journal of bacteriology.

[29]  H. Boyer,et al.  A complementation analysis of the restriction and modification of DNA in Escherichia coli. , 1969, Journal of molecular biology.

[30]  S. Falkow,et al.  Polynucleotide Sequence Relationships among Members of Enterobacteriaceae , 1969, Journal of bacteriology.