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Genetics and Molecular Biology |
Department of Microbiology and Immunology, and Division of Clinical Microbiology, University of Western Ontario, London, Ontario, N6A 5C1, Canada1
Instituto de Investigaciones Bioquímicas, Fundación Campomar, Antonio Machado 151, 1405 Buenos Aires, Argentina2
Author for correspondence: Miguel A. Valvano. Tel: +1 519 661 3996. Fax: +1 519 661 3499. e-mail: mvalvano{at}julian.uwo.ca
In previous studies the authors cloned and characterized the DNA sequence of the regions at both ends of the O7-specific lipopolysaccharide (LPS) biosynthesis cluster of Escherichia coli VW187 (O7:K1), and identified the biosynthetic genes for dTDP-rhamnose and GDP-mannose, as well as one of the candidate glycosyltransferases. In this work the complete DNA sequence of a 6·9 kb intervening region is presented. Seven new ORFs were identified. All the functions required for the synthesis and transfer of the O7 LPS were assigned on the basis of complementation experiments of transposon insertion mutants, and amino acid sequence homology to proteins involved in LPS synthesis of other bacteria. Of the seven ORFs, two encoded membrane proteins that were homologous to the O-antigen translocase (Wzx) and polymerase (Wxy), two were involved in the biosynthesis of dTDP-N-acetylviosamine, and the remaining three showed homologies to sugar transferases. The O antigen chain length regulator gene wzz was also identified in the vicinity of the O7 polysaccharide cluster. O7-specific DNA primers were designed and tested for serotyping of O7 E. coli strains.
Keywords: Escherichia coli, wb (rfb) gene cluster, lipopolysaccharide (LPS), O antigen, PCR serotyping
Abbreviations: ECA, enterobacterial common antigen; FucN, 4-amino-4,6-dideoxy-D-galactose; FucNac, 4-acetamido-4,6-dideoxy-D-galactose; GlcNAc, N-acetylglucosamine; Vio, viosamine, 4-amino-4,6-dideoxyglucose; VioNAc, N-acetylviosamine (4-acetamido-4,6-dideoxyglucose)
The GenBank accession number for the DNA sequence reported in this paper is AF125322.
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