FIGURE 5.

Effect of 3HPP and DHPP on the transcriptional activation from Pa promoter mediated by MhpR-His. A, single round in vitro transcription was carried out by using the plasmid pJCDAR as template. The concentrations of proteins were 100 nm RNAp, 100 nm MhpR-His, and 200 nm CRP. Lanes 1–6 contained 1, 10, 50, 100, 500, and 1000 μm 3HPP, respectively. Lane 7 contained 1 mm PP. Lanes 8–13 contained the same gradient but in the presence of 1 mm PP. Lanes 14–19 contained 1, 10, 50, 100, 500, and 1000 μm DHPP, respectively. Lanes 20–25 contained the same gradient but in presence of 1 mm PP. Arrows point to the Pa-derived mRNA (181 nucleotides) and the vector-derived RNA1 (108–109 nucleotides). B, Pa transcription levels (in arbitrary units) produced from pJCDAR template in the presence of 1–1000 μm 3HPP or DHPP in the absence and presence of 1 mm PP.