FIG. 3.

Spo0A(A257V)∼P stimulates spoIIG transcription in vitro. Phosphorylated or unphosphorylated Spo0A or Spo0A(A257V) was incubated with a linear DNA fragment encoding the spoIIG operon promoter and the initiating nucleotides ATP and GTP. RNAPσ^A was added to the mixture and allowed to initiate transcription for 2 min prior to addition of the remaining nucleotides and heparin. Following a 5-minute incubation, the elongated transcripts were separated by electrophoresis through an 8% denaturing polyacrylamide gel. ³²P-labeled spoIIG transcripts were detected by exposure to a phosphor screen (A), and the level of transcription was determined by PhosphorImager analysis (B). Filled circles, Spo0A∼P; filled triangles, Spo0A; open circles, Spo0A(A257V)∼P; open triangles, Spo0A(A257V). WT, wild type. A representative phosphorimage is shown. The values reflect the average of three independent experiments. The error bars indicate standard deviations.