Cloning and sequence of the genes downstream of the wbf gene cluster of Vibrio cholerae serogroup O139 and analysis of the junction genes in other serogroups

Abstract

The DNA sequence of the O-antigen biosynthesis cluster (wbf) of a recently emergent pathogen, Vibrio cholerae serogroup O139, has been determined. Here we report the sequence of the genes downstream of the O139 wbfX gene and analysis of the genes flanking the wbf gene cluster in other serogroups. The gene downstream of wbfX, designatedrjg (right junction gene), is predicted to be not required for O-antigen biosynthesis but appears to be a hot spot for DNA rearrangements. Several variants of the rjg gene (three different insertions and a deletion) have been found in other serogroups. DNA dot blot analysis of 106 V. choleraestrains showed the presence of the left and right junction genes,gmhD and rjg, respectively, in all strains. Further, these genes mapped to a single I-CeuI fragment in all 21 strains analyzed by pulsed-field gel electrophoresis, indicating a close linkage. The insertion sequence element IS1358, found in both O1 and O139 wb regions, is present in 61% of the strains tested; interestingly, where present, it is predominantly linked to the wb region. These results indicated a cassette-like organization of the wb region, with the conserved genes (gmhD and rjg) flanking the divergent, serogroup-specific wb genes and IS1358. A similar organization of the wb region in other serogroups raises the possibility of the emergence of new pathogens by homologous recombination via the junction genes.