FIG. 2.

Q-RT-PCR analysis of pilE transcript levels. (A) The strains were grown on solid medium with the indicated level of IPTG. Total RNA was isolated with and reverse transcribed with specific primers. Transcript copies were determined by FRET hybridization probe detection in a Roche LightCycler compared to known amounts of cloned DNA standards. Values are the percentage of the wild-type RM11.2 strain value. Shown is an average of three replicates. (B) The strains were grown in liquid medium with the indicated level of IPTG. Q-RT-PCR was performed as described for panel A using por primers and probes, and RNA samples were normalized to contain similar levels of por transcript levels (data not shown). pilE transcript numbers for fully induced and uninduced regulatable (reg) pilE variants were determined and expressed as the percentage of wild-type (recA6) pilE transcript level for each variant.