FIG 3.

Electrophoretic mobility shift assays using purified LacI transcriptional regulators with DNA fragments from the promoters of regulated genes. Approximately 0.03 μM, 0.02 μM, and 0.03 μM DNA fragments from the upstream genomic regions of Clo1313_2022, Clo1313_1398, and Clo1313_0395 were incubated with increasing amounts of purified protein encoded by Clo1313_2023, Clo1313_0089, and Clo1313_0396, respectively. Lanes were loaded as follows: lane 1, 100-bp marker (Life Technologies); lane 2, DNA only; lanes 3 to 11, DNA plus LacI regulator (for GlyR1 and GlyR2, 0.016, 0.03, 0.06, 0.13, 0.25, 0.50, 0.75, 1.0, 1.5 μM, respectively; for GlyR3, 0.06, 0.13, 0.25, 0.50, 0.75, 1.0, 1.5, 2.0, 2.5 μM); lane 12, protein only. FP indicates where the free probe is running on the gel.