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. 2018 Mar 23;19(8):1859–1872. doi: 10.1111/mpp.12664

Figure 6.

Figure 6

Clp can bind to the promoter region of PXO_03177. (A) The direct physical interaction between Clp and the PXO_03177 promoter region was detected in Escherichia coli. Positive control, co‐transformant containing pBX‐R2031 and pTRG‐R3133; negative control, co‐transformant containing pBXcmT‐PXO_03177 and empty pTRG; pTClp/p‐PXO_03177, co‐transformant possessing both pTRG‐Clp and pBXcmT‐PXO_03177; −3AT‐Strr, no selective Luria–Bertani (LB) medium plate; +3AT+Strr, M9‐based selective medium plate. (B) Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) analysis of the purified glutathione transferase (GST)‐tagged Clp. (C) The probe and mutational probe sequence of PXO_03177. The putative Clp binding site and its mutations are underlined. (D) Clp binds to the PXO_03177 promoter (P‐PXO_03177) in vitro. Unlabelled PXO_03177 probe (code probe) was found to competitively inhibit the formation of the labelled Clp–DNA complex. Arrows indicate free DNA and Clp–DNA complexes. No binding of Clp with the mutational probe (Pm‐PXO_03177) was observed.