FIG. 3.

Gel mobility shift assays with purified response regulator LiaR. For the reactions, LiaR was phosphorylated by acetyl phosphate (ac-P), with the exception of the assay whose results are shown in the lower left panel, as described in Materials and Methods. PCR products of P_liaI (180 bp), P_yhcY, and P_ydhE were incubated with an increasing molar excess of phosphorylated LiaR (lane 1, 50:1; lane 2, 75:1; lane 3, 100:1) followed by gel electrophoresis on native 8% polyacrylamide gels and detection of the DNA with SYBR green I. The first lane in each panel corresponds to free DNA (−). An internal fragment of bceA (161 bp) was always included as a negative control (C).