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Fig. S1. Location of chxA within the V. cholerae genome and approximate locations of selected primer sequences. The chxA gene exists as a single gene indel in V. cholerae strains shown here. Approximate locations of conserved primers for future studies of chxA prevalence (ChxAF2 and ChxAR1), and primers for determining its chromosomal location (VC1645_F and ChxAR1; VC1644_F and ChxAF3) are indicated by arrows.

Fig. S2.chxA alleles are located adjacent to hypothetical gene VC1644 in V. cholerae strains carrying chxA genes that fall into clade 1 and clade 2. PCR primers VC1644_F and ChxAF3 (Table S2) were used amplify 2.1 kb from VC1644 to chxA. From left to right, after the ladder, amplicons represent strains M1, M45, B31, B55, C7, C39, C160, M6, M33, B33, B54, C62, C74, SIO, TP and N16961.

Fig. S3. Variation in regions upstream of the chxA alleles falling in clade 2. PCR primers VC1645_F and ChxAR1 (Table S2) were used amplify 1.5 kb from VC1645, a predicted hydrolase, to chxA. From left to right, after the ladder, amplicons represent strains M1, M45, B31, B55, C7, C39, C160, M6, M33, B33, B54, C62, C74, SIO, TP and N16961. Amplification of a 1.5 kb fragment was unsuccessful in the strains with chxA alleles falling in clade 2: M6, M33, B33 and B54.

Table S1. Strains used in this study.

Table S2. Primers used in this study.

Appendix S1. Supplementary experimental procedures.

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Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.