Figure 6
HpaR1 enhances the transcription of gumB promoter in vitro.
(A) RNA was generated from a 687-bp template DNA fragment extending from −345 to +342 relative to the transcriptional initiation site (TIS) of the gumB promoter using E. coli RNA polymerase (RNAP) holoenzyme. (B) A template DNA fragment containing the hrpG promoter was used as a control. (C) A 329-bp template DNA fragment extending from +13 to +342 relative to the TIS of gumB promoter was also used as a control. 2 nM template DNA was incubated with a series of amounts of His6-HpaR1 protein before beginning transcription by the addition of 0.05 U of RNAP. Transcription products were then run on a 5% denatured polyacrylamide gel containing 7 M urea in 1× TBE electrophoresis buffer. Lane 0, template DNA alone; Lane 1, template DNA with RANP; Lanes 2–4, template DNA with RANP and 10, 15 and 20 nM His6-HpaR1.
