FIG. 4.

In vitro transcription from the codBA promoter. The DNA template was supercoiled plasmid pCB592. The reactions in lanes 1 through 6 contained purified RNA polymerase from E. coli. Lane 1 contained no NAC. Lanes 2 through 4 contained 0.1, 0.2, and 1.0 μg of NAC derived from K. pneumoniae, respectively. Lanes 5 and 6 contained 1 μg of NAC (from K. pneumoniae and E. coli, respectively) prepared as an MBP-NAC fusion protein and cleaved with factor Xa to release active NAC immediately before addition to the reaction mixture. Lane 7 contained labeled DNA size standards of 210, 245, 400, and 510 bp. The arrow indicates the runoff transcript from the codB promoter.