FIG 5.
The putative transcription regulator Rv0634A does not regulate the tha operon. (A) Genetic map showing the rv0634A and sh ble allelic exchange. Primers for the deletion analysis by PCR are symbolized by arrows. (B and C) PCR products with the a+b and a+c primer pairs to confirm the deletion of the rv0634A gene. Lysates of a Δrv0634A strain (1) and of a WT clone (2) were used as PCR templates. (D) Monitoring by RT-qPCR of the expression (log2) of the rpmG2, hadA, and rv0634A genes relative to the WT strain values (WT/pLAM) in the rv0634A mutant (Δrv0634A/pLAM) and in the rv0634A-overexpressing background (Δrv0634A/pLAM-rv0634A). Each strain was cultivated in the presence of 0.2% acetamide. Error bars show standard errors of the means (SEM) from three biological triplicates. (E) Expression, in relative fluorescent units (RFU), of rthp::gfp (FA) and thap3::gfp (FB) fusions in M. smegmatis carrying a plasmid expressing rv0634A from an acetamide-inducible promoter (pLAM12-rv0634A) or not (pLAM12). Each strain was cultivated in either the presence (+) or absence (−) of 0.2% acetamide. pGBG was the gfp promoterless control.