Fig 5.

Identification of the GenR binding sites upstream of genDFM, genR, and genKH operons by DNase I footprinting analysis. The reaction mixtures contained approximately 200 ng of end-labeled PCR products. These were amplified with prnagRD02 primer and ³²P-labeled pegenDFM01 primer (A), pgenR01 and ³²P-labeled pegenR01 (B), or pgenKH02 and ³²P-labeled pegenKH04 (C). Before DNase I treatment, labeled DNA was preincubated with His₆-GenR for 30 min in the presence of 300 nM 3-HBA. Standards were generated by sequencing with ³²P-labeled primers pegenDFM01, pegenR01, and pegenKH04, respectively. The concentrations (μM) of purified His₆-GenR and the GenR-protected sequences are indicated. The nucleotide sequence around the TSS (+1) is shown by solid arrows, and the GenR binding sites are represented by brackets. Arrows indicate the direction of transcription.