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. 2017 May 16;8:869. doi: 10.3389/fmicb.2017.00869

FIGURE 9.

FIGURE 9

Electrophoretic mobility shift assay (EMSA) using CRP and upstream regions of lldP. (A) Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) of N-his-CRP protein samples. Protein samples (5 μg) were analyzed on 12.5% SDS-polyacrylamide gels. Lane 1, Escherichia coli BL21 (DE3) (pET-crp) crude extract; lane 2, purified N-his-CRP; lane 3, molecular weight marker. (B) DNA fragments used as probes. Positions of 5′ and 3′ ends of the fragments relative to TSSlldP (+1) are shown. White boxes represent the putative CRP-binding motif. The mutated sequences in PBlldP3m are shown in bold. (C) Binding of CRP to each probe. DNA-binding reactions were performed in the presence (+) or absence (–) of CRP, cAMP, and specific competitor (2 μM unlabeled PBlldP3 probe).