Figure 5

Binding of His-PhoP and His-PhoP _N149 to P _pmrD . (A) Specific binding of recombinant His-PhoP protein to the putative pmrD promoter. EMSA was performed by using the ³²P-labeled DNA probe of P _pmrD incubated with increasing amounts of the His-PhoP (lanes 2 to 5), with 40 pmole of His-PhoP plus increasing amounts of the unlabeled P _pmrD DNA (specific competitor, lane 6 to 9), or with excess amounts of non-specific competitor DNA (lane 10 and 11). The amounts of recombinant proteins and DNA probes used are indicated in the figure. (B) EMSA was performed with 0, 4 or 40 pmole of His-PhoP (lanes 1 to 3), His-PhoP_N149 (lanes 4 to 6) or 100 pmole of BSA (lane 7). The arrows indicate the PhoP/P _pmrD complex and free probe of P _pmrD .