FIGURE 6.
cAMP-CRP binding directly to tolB and rcsF promoter was identified by EMSA and lacZ fusion assay. (A) The tolB-pal-cpoB operon and the promoter sequence. The large arrows represent the open-reading frames, and the CRP binding site was marked as red color and underlined. (B) The rcsF-trmO operon and the promoter sequence. (C) LacZ fusion assay of the cAMP-CRP binding to the tolB and ftsI promoter regions. Recombinant pHRP309 plasmids were transferred into the WT, crp-knockout, and crp complementary mutant of K. pneumoniae. Error bars represent standard deviation. ∗∗P < 0.01; ∗∗∗P < 0.001; n.s., not significant; comparison was performed by one-way ANOVA. (D) cAMP-CRP binding directly to tolB and rcsF promoter regions was identified by EMSA. Ten nanogram unlabeled probe (cold probe) was added to compete with labeled probe for binding His6-CRP protein.