1887

Abstract

Until the discovery of the epeat (VCR), the gene capture and expression systems termed integrons had been typically associated with antibiotic-resistance gene cassettes with usually less than five genes in an array. A method is described for the cloning of the ends of large cassette arrays. Conserved restriction sites within VCRs facilitated the mapping by Southern hybridization and cloning of the 5’ end of the VCR array, and using appropriate fragments it was possible to develop a physical map of the region of the chromosome. Sequence determination of the predicted beginning of this region revealed , a member of the integron family of integrases. Comparison of these sequences from El Tor, Classical and serotype O134 strains identified the 3’ end of the site, thereby defining the class 4 integron in one of the chromosomes, and providing the first evidence for integron-like site-specific recombination within . Conduction assays demonstrated IntI1-mediated recombination between VCRs. Restriction mapping places the sequences of and 26 VCR gene cassettes in arrays within a 120 kb region of the O1 strain 569B genome. This region contains an estimated 150 VCR gene cassettes, dwarfing previously described arrays. Southern analysis of genomic DNA from strains of , and a number of serotypes revealed fragments that hybridized with VCR-specific probes but showed a high degree of restriction fragment length polymorphism. These data facilitate the identification of part of a new class 5 integron from .

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2000-10-01
2019-10-18
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