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Distribution of IS1358 and linkage to rfb-related genes in Vibrio anguillarum

Microbial Pathogenesis Unit, Department of Microbiology and Immunology, The University of Adelaide, Adelaide, SA 5005, Australia¹
Mikrobiologie II, Universität Tübingen, D-72076 Tübingen, Germany²
Microbiology and Molecular Biology, AstraZeneca R&D Boston, 128 Sidney Street, Cambridge, MA 02139-4239, USA³

Author for correspondence: Paul A. Manning. Tel: +1 617 576 3900. Fax: +1 617 576 3030. e-mail: paul.manning{at}astrazeneca.com

The insertion sequence IS1358 is linked to the rfb regions of both Vibrio cholerae O1 and O139, and its location was suggestive of a role in generating new combinations of rfb genes. This provoked an examination of the distribution and localization of IS1358 in Vibrio anguillarum. IS1358 was widely distributed in a number of V. anguillarum serogroups. In particular, when cosmid clones of V. anguillarum O1 were screened with IS1358 and subsequently subcloned and sequenced, it was found that rfb-like genes were linked to this region. Furthermore, when the previously identified genes virA and virB from V. anguillarum O1, now known to be involved in LPS biosynthesis, were used as probes, it was discovered that they too are present on the same large EcoRI fragment as IS1358. This clearly indicated that IS1358 was linked to the rfb region of V. anguillarum O1. Further analysis of the location of IS1358 in other serotypes indicated that V. anguillarum O2 also has IS1358 associated with rfb-like genes. In V. anguillarum O2 there is more than one copy of IS1358, suggesting that this element is a site for recombination, gene duplication or that it may be capable of transposition. Following this latter premise, IS1358 elements from a variety of V. anguillarum strains have been cloned and sequenced. Only those strains with multiple copies of IS1358 produce a full-length putative transposase, as shown by protein overexpression, further strengthening the argument that the element is transposing within these strains.

Keywords: O-antigen, polysaccharide, insertion sequences, transposase, evolution

The GenBank accession numbers for the IS1358 sequences are U93587–U93597.

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