FIGURE 7.
Effect of BzdR or BzdRΔL and benzoyl-CoA on in vitro transcription from PN. Multiple round in vitro transcription reactions were performed as indicated under “Experimental Procedures” by using pJCD-PN, a plasmid template that produces a 184-nucleotide mRNAs from PN, 50 nm E. coli RNA polymerase, and 20 nm Fnr* activator. The transcription reactions were carried out in the absence of repressor (lane 2) or in the presence of 25 nm (lanes 3 and 4), 50 nm (lane 5), or 100 nm (lane 6) of purified His6-BzdR or 25 nm (lanes 7 and 8), 50 nm (lane 9), or 100 nm (lane 10) of purified His6-BzdRΔL. Benzoyl-CoA was added at 1 mm (lanes 4–6 and 8–10). Lane 1, control assay without RNA polymerase.