FIG. 6.

Binding and transcription with recombinant HrcA at an elevated temperature. (A) EMSA reactions with 0.5 nM ³²P-labeled dnaK CIRCE probe and various concentrations of recombinant HrcA were performed in triplicate, and the results were quantified by phosphorimager analysis. The reaction mixtures were preincubated at room temperature for 15 min and then shifted to 37°C (▪), 42°C (▴), or 56°C (♦) for 5 min. The error bars indicate standard deviations from the mean. (B) In vitro transcription with E. coli RNA polymerase. The upper band was from the dnaK promoter, and the lower band was from the omcB promoter. The lanes show (from left to right) transcription at 37°C with no HrcA, transcription at 37°C with 1,150 nM HrcA, transcription at 42°C with no HrcA, and transcription at 42°C with 1,150 nM HrcA.