Figure 5

Footprint analysis of the sacB promoter by using His-DegU. A. Preparation of the probes and the sequencing ladders was the same as that in figure 4. The probes (40 nM) were incubated with increasing amounts of His-tagged DegU (0.3 and 0.6 μM) and subjected to DNase I cleavage. For footprint assays, we used the procedures developed for biotinylated DNA probes as described previously [47]. The sequencing ladder is shown in lanes G, A, T and C. The nucleotide sequence of the promoter region is shown. The brackets and arrowheads indicate the protected regions and hypersensitive sites, respectively. B. Arrows above the sequence indicate the DegU-binding motifs. The numbers indicate the nucleotide positions relative to the transcription start site.