FIG 8.
Characterization of the core and regulatory elements of Pgan. (A) Identification of the transcription start site of Pgan was carried out using primer extension reactions. The generated cDNA probes (P1 and P2 repeats) were compared with the sequencing reaction using the dideoxy chain-termination method (A, C, G, T). (B and C) The DNase I footprinting reactions for the Pgan coding strand DNA (B) and noncoding strand DNA (C) were compared to the dideoxy chain-termination reactions (A, C, G, and T). The DNase I footprinting reactions were performed without GanR as the negative control (−) or with different amounts of GanR (1.95, 3.90, and 7.79 μM). The bar shows the protected DNA region in each DNase I footprinting reaction. (D) The DNA sequence of the Pgan region. The promoter core elements (−35 box and extended −10 box) and the transcription start site (+1) are shown by rectangles. The protected DNA region is shown by solid lines. The arrow shows the start codon of ganS. The predicted cre site is highlighted in gray.