Fig 7.

Mutational analyses of the four GenR binding sites. (A to D) EMSAs using the wild-type and mutated DNA fragments. Probes DFMan01, DFMan02, R-KHan01, and R-KHan02 contained the intact GenR binding sites DFMn01, DFMn02, R-KHn01, and R-KHn02, respectively. Probes DFMan01m, DFMan02m, R-KHan01m, and R-KHan02m contained the mutated sites described in Materials and Methods. The amounts (nM) of GenR used in lanes 1 to 5 are indicated. The free probes are indicated by open arrows, and the retarded DNA fragments are indicated by solid arrows. (E) β-Galactosidase activity driven by genDFM, genR, and genKH promoters with GenR binding sites in strain RES167. DFMa, Ra, and KHa are the wild-type GenR binding sites. DFMan01m, DFMan02m, DFMan12m, Ran02m, and KHan02m are the mutated binding sites. The β-galactosidase activity analyses were performed as described in the text. The data are derived from at least three independent measurements, and error bars indicate standard deviations.