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Identification and characterization of preferred DNA-binding sites for the Thermus thermophilus transcriptional regulator FadR

Fig 4

Quantitative EMSA analysis of FadR-binding to consensus and mutant sequences.

Shown are LICOR images of IRD700-labeled FadR consensus or point-mutated DNAs, as indicated, incubated with a twofold (wt, m1, m3, m4, m5, m7) or tenfold (m2, m6) titration of FadR protein, as indicated. (S) FadR-DNA complex, (T) uncomplexed DNA. (A) ST2_FadR_R5_wt consensus DNA; 0, 0.038, 0.075, 0.15, 0.3, 0.6, 1.2 nM FadR. (B) ST2_FadR_R5_m1 mutant DNA; 0, 0.38, 0.75, 1.5, 3.0, 6.0, 12 nM FadR. (C) ST2_FadR_R5_m2 mutant DNA; 0, 0.06, 0.6, 6, 60, or 600 nM FadR. (D) ST2_FadR_R5_m3 mutant DNA; 0, 3.8, 7.5, 15, 30, 60, 120 nM FadR. (E) ST2_FadR_R5_m4 mutant DNA; 0, 0.75, 1.5, 3.0, 6.0, 12, 24 nM FadR. (F) ST2_FadR_R5_m5 mutant DNA; 0, 7.5, 15, 30, 60, 120, 240 nM FadR. (G) ST2_FadR_R5_m6 mutant DNA; 0, 0.06, 0.6, 6, 60, or 600 nM FadR. (H) ST2_FadR_R5_m7 mutant DNA; 0, 7.5, 15, 30, 60, 120, 240 nM FadR. Binding site sequence and KD values are indicated below each panel. Lowercase nucleotides indicate mutation from consensus FadR sequence.

Fig 4

doi: https://doi.org/10.1371/journal.pone.0184796.g004