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. Author manuscript; available in PMC: 2018 May 2.
Published in final edited form as: Biochemistry. 2017 Apr 21;56(17):2315–2327. doi: 10.1021/acs.biochem.7b00103

Figure 5.

Figure 5

BifR binds specifically to the emrB-bif R promoter DNA. (A) EMSA showing 355 bp operator DNA (0.8 nM) titrated with increasing concentration of BifR (lane 2, 0.5 nM; lane 3, 1 nM; lane 4, 2.5 nM; lane 5, 5 nM; lane 6, 8 nM; lane 7, 10 nM; lane 8, 25 nM; lane 9, 50 nM; lane 10, 100 nM; lane 11, 150 nM; lane 12, 250 nM); reaction in lane 1 contains DNA only. Free DNA is identified as F and complexes are identified as C1 and C2 at the right. (B) EMSA showing 355 bp operator DNA (0.8 nM) titrated with increasing concentration of BifR-C104A (lane 2, 0.5 nM; lane 3, 1 nM; lane 4, 2.5 nM; lane 5, 5 nM; lane 6, 8 nM; lane 7, 10 nM; lane 8, 25 nM; lane 9, 50 nM; lane 10, 100 nM; lane 11, 150 nM; lane 12, 250 nM). (C) BifR (1.3 nM) bound to labeled operator DNA challenged with unlabeled 355 bp operator DNA (0.8–45 nM, lanes 3–8) or equivalent concentration of nonspecific DNA pET28b (lanes 10–15). Reaction in lane 1 contains free DNA; reactions in lanes 2 and 9 contain no competitor DNA. (D) Fractional complex plotted as a function of BifR (●, dashed line) and BifR-C104A (■, solid line) concentration. Error bars represent the standard deviation of three independent repeats. (E) BifR-DNA complexes titrated with increasing concentration of salicylate (1–3 mM; lanes 3–5). Reaction in lane 2 contains no inducer. Reaction in lane 1 contains DNA only.