Fig 3.
Binding of RamA, GntR1, GntR2, and IolR to the pck promoter region. (A) DNA fragments (Ppck 1 to Ppck 13) used to determine the location of the RamA, GntR1, GntR2, and IolR binding site(s) in the pck promoter are shown. The transcription start site (TSS) of the pck gene is indicated by an arrow labeled +1. Black bars indicate the DNA fragments used for electrophoretic mobility shift assays (EMSAs). The numbers given to the right show the positions of the fragment ends relative to the TSS. The GlxR, RamA, GntR1/GntR2, and IolR binding sites are indicated with checkered, white, black, and striped boxes, respectively. (B) Results of EMSAs using different fragments of the pck promoter region and His-tagged RamA, GntR1, GntR2, and IolR. The DNA fragments used (Ppck 1 to Ppck 13) are indicated at the top of each gel. Free DNA and DNA-protein complexes are indicated with white and black arrowheads, respectively. Nonspecific bands are indicated with asterisks.