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In order to identify additional adherence factors of EHEC, we generated transposon mutants of a clinical EHEC isolate of serotype O111:H\u2013, which displayed high levels of adherence to cultured Chinese hamster ovary (CHO) cells. One mutant was markedly deficient in CHO cell adherence, human red blood cell agglutination and autoaggregation. Sequence analysis of the gene disrupted in this mutant revealed a 9669\u2003bp novel chromosomal open reading frame (ORF), which was designated <jats:italic>efa1<\/jats:italic>, for <jats:styled-content>E<\/jats:styled-content>HEC <jats:styled-content>f<\/jats:styled-content>actor for <jats:styled-content>a<\/jats:styled-content>dherence. <jats:italic>efa1<\/jats:italic> displayed 28% amino acid identity with the predicted product of a recently described ORF from the haemolysin\u2010encoding plasmid of EHEC O157:H7. The amino termini of the putative products of these two genes exhibit up to 38% amino acid similarity to <jats:italic>Clostridium difficile<\/jats:italic> toxins A and B. <jats:italic>efa1<\/jats:italic> occurred within a novel genetic locus, at least 15\u2003kb in length, which featured a low G+C content, several insertion sequence homologues and a homologue of the <jats:italic>Shigella flexneri<\/jats:italic> enterotoxin ShET2. DNA probes prepared from different regions of <jats:italic>efa1<\/jats:italic> hybridized with all of 116 strains of attaching\u2013effacing <jats:italic>E. coli<\/jats:italic> (AEEC) of a variety of serotypes, including enteropathogenic <jats:italic>E. coli<\/jats:italic> (EPEC) and EHEC, but with none of 91 non\u2010AEEC strains. Nevertheless, <jats:italic>efa1<\/jats:italic> was not required for the attachment\u2013effacement phenotype, and the <jats:italic>efa1<\/jats:italic> locus was not physically linked to the locus for enterocyte effacement (LEE) pathogenicity island, which is responsible for this phenotype in EPEC. These findings suggest that <jats:italic>efa1<\/jats:italic> encodes a novel virulence\u2010associated determinant of AEEC, which contributes to the adhesive capacity of these bacteria.<\/jats:p>","DOI":"10.1046\/j.1365-2958.2000.01690.x","type":"journal-article","created":{"date-parts":[[2003,3,12]],"date-time":"2003-03-12T11:28:31Z","timestamp":1047468511000},"page":"275-288","source":"Crossref","is-referenced-by-count":204,"title":["Identification of a novel genetic locus that is required for <i>in vitro<\/i> adhesion of a clinical isolate of enterohaemorrhagic <i>Escherichia coli<\/i> to epithelial cells"],"prefix":"10.1111","volume":"35","author":[{"given":"Larissa","family":"Nicholls","sequence":"first","affiliation":[]},{"given":"Travis H.","family":"Grant","sequence":"additional","affiliation":[]},{"given":"Roy M.","family":"Robins\u2010Browne","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[2002,3]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.39.1.315-335.1983"},{"key":"e_1_2_6_3_2","unstructured":"Ausubel F.M. 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