Figure 7.

The role of core and auxiliary motifs in Mgal-WhiA repressor function. (A) The overview of the transposon vector used to test promoter activity. The translation of the transposase gene in E. coli is blocked by three nonsense codons, TGA, that are readthrough in mycoplasmas as Trp-codons. The sequence transferred to the genome is encompassed between OIR and IIR transposition motifs and integrates randomly. (B) Transcription activity of WT rpsJ promoter and rpsJ promoter with substitutions in core or auxiliary (aux) sequences. The level of rpsJ mRNA was measured using RT-qPCR relatively to enolase (eno) transcript. The substitutions in either core or aux motifs induce promoter activation relative to WT sequence. Bars indicate standard error. (C) The sequences of promoters used for testing. Mgal-WhiA binding site is highlighted in bold, substitutions are highlighted in red. TSS was identified in Mazin et al. (2014).