Characterization of C. difficile R20291::sinR’.

(A)C. difficile cultures were grown in 70:30 medium for 30 h under anaerobic conditions and Sporulation frequency (CFU/ml of ethanol resistant spores) of R20291, sinR’ mutant was determined. The data shown are means ± standard errors of three biological replicates. (B) Motility assays of the C. difficile R20291, sinR’ mutant and complemented sinR’ mutant. The experiments were repeated at least three times independently (*, P≤0.05 by a two-tailed Student's t-test). (C) Toxin production measured by ELISA. Statistical analysis was performed using one way-ANOVA with Dunnett’s multiple comparisons test comparing values to the average of the parent with vector control (***<0.0005, *< 0.05 p-value).