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Subtractive hybridization and identification of putative adhesins in a Shiga toxin-producing eae-negative Escherichia coli.
Vidal, Maricel; Prado, Valeria; Whitlock, Gregory C; Solari, Aldo; Torres, Alfredo G; Vidal, Roberto M.
Afiliación
  • Vidal M; Programa de Microbiología y Micologia, ICBM, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
  • Prado V; Programa de Microbiología y Micologia, ICBM, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
  • Whitlock GC; Department of Clinical Laboratory Sciences, University of Texas Medical Branch, Galveston, TX 77555-1070, USA.
  • Solari A; Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555-1070, USA.
  • Torres AG; Programa de Biologia Celular y Molecular, ICBM, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
  • Vidal RM; Department of Pathology and Sealy Center for Vaccine Development, University of Texas Medical Branch, Galveston, TX 77555-1070, USA.
Microbiology (Reading) ; 154(Pt 12): 3639-3648, 2008 Dec.
Article en En | MEDLINE | ID: mdl-19047731
Adherence to epithelial cells by specific adhesins is a characteristic of Shiga toxin-producing Escherichia coli (STEC) strains. The eae-encoded protein intimin is the main adhesin implicated in intestinal colonization in vivo. We recently showed that STEC strains isolated in Chile displayed a wide variety of adhesins; here we demonstrate that some of these STEC strains are eae-negative and still adhere to epithelial cells at a level 100-fold higher than enterohaemorrhagic E. coli (EHEC) O157 : H7 prototype strain EDL933. This phenotype is associated with the presence of adherence factors different from the intimin protein. Subtractive hybridization between EHEC EDL933 and STEC eae-negative strain 472-1 was used to identify regions implicated in adhesion. In addition to the saa gene, we identified 18 specific genes in STEC 472-1, 16 of which had nucleotide identity to Salmonella ST46 phage genes; the two remaining ones shared identity to a gene encoding a hypothetical protein of uropathogenic E. coli. The DNA sequence of the STEC 472-1 psu-int region identified five open reading frames with homology to phage genes. We constructed mutant strains in the saa gene and the psu-int region to study the participation of these genes in the adherence to epithelial cells and our results demonstrated that STECDeltasaa and STECDeltapsu-int mutants displayed a 10-fold decrease in adherence as compared to the STEC 472-1 wild-type strain. Overall, our results suggest that STEC strain 472-1 adheres to epithelial cells in an eae-independent matter and that saa and psu-int participate in this adhesion process.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Adhesinas de Escherichia coli / Adhesinas Bacterianas / Proteínas de Escherichia coli / Escherichia coli Shiga-Toxigénica / Hibridación de Ácido Nucleico Tipo de estudio: Diagnostic_studies / Prognostic_studies Límite: Humans Idioma: En Revista: Microbiology (Reading) Asunto de la revista: MICROBIOLOGIA Año: 2008 Tipo del documento: Article País de afiliación: Chile Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Adhesinas de Escherichia coli / Adhesinas Bacterianas / Proteínas de Escherichia coli / Escherichia coli Shiga-Toxigénica / Hibridación de Ácido Nucleico Tipo de estudio: Diagnostic_studies / Prognostic_studies Límite: Humans Idioma: En Revista: Microbiology (Reading) Asunto de la revista: MICROBIOLOGIA Año: 2008 Tipo del documento: Article País de afiliación: Chile Pais de publicación: Reino Unido