Fig 3. EMSA demonstrating the interaction of GlnR and PmrA to the promoters of genes in the GlnR regulon.
The pglnR-, pglnQ-, pgdhA-, pcitB-, pnrgA-1-, pnrgA-2-, pglnP- and ppmrA-specific DNA probes were mixed with various amounts of protein extracts. All reactions were performed with 0.025 pmol of biotin-labeled probes. (A) Lane 1, probe only. Lanes 2 to 7, use of 8 to 256 nM of MBP-GlnR in the reactions in twofold increments, respectively. Lane 8, use of 256 nM of MBP-GlnR with a non-specific competitor. In the reactions containing the ppmrA probe, 16 nM to 512 nM of MBP-GlnR was used instead. (B) Lane 1, probe only. Lanes 2 to 8, use of 8 nM to 512 nM of MBP-PmrA in the reactions in twofold increments, respectively. Lane 9, use of 512 nM of MBP-PmrA with a non-specific competitor.