FIG. 8.

DNA binding assays. Gel mobility shift analysis with the orf10-orf11 intergenic region was performed as described in Materials and Methods, using the 316-bp EspI-AvaI fragment as the probe. Lanes: 1 and 12, without protein addition; 2 and 3, with crude extracts from cultures of E. coli K12ΔH1Δtrp carrying the control plasmid pAZe3ss, grown at 30 and 42°C, respectively; 4 and 5, with crude extracts from cultures of the same strain harboring plasmid pCNB019, grown at 30 and 42°C, respectively; 6 to 9, with 1.2, 2.5, 12.5 and 25 ng of the purified S. coelicolor Orf10 protein, respectively; 10, with 25 ng of Orf10 and fivefold molar excess of unlabeled orf10-orf11 intergenic region; 11, with 25 ng of Orf10 previously treated for 15 min at 100°C.