FIG. 5.

In vitro transcription assay. Single-round transcription in vitro of 0.1 pmol ung DNA template (−452 to +86 of promoter region) (A, lanes 5 to 8; B, lanes 5 to 8) or lacUV5 DNA template (A, lanes 1 to 4; B, lanes 1 to 4) was performed in the presence (B) or absence (A) of 10 mM acetylphosphate. The amounts of CpxR were as follows: lanes 1 and 5, 0 pmol; lanes 2 and 6, 1.25 pmol; lanes 3 and 7, 2.5 pmol; lanes 4 and 8, 5 pmol. Electrophoresis was performed with a 6% polyacrylamide sequencing gel. Bold arrows indicate the ung and lacUV5 transcripts. (C) ung (circles) and lacUV5 (square) transcripts in the absence (open symbols) and presence (closed symbols) of CpxR were quantified with BAS 1000 Mac (Fuji film). The relative value in shown as a ratio between each transcript and that in the absence of CpxR.