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. 2018 Mar 27;9:599. doi: 10.3389/fmicb.2018.00599

FIGURE 5.

FIGURE 5

Electrophoretic mobility shift assays (EMSAs) of the interaction between LhrC4 and lmo0484 mRNA. (A) LhrC4 mutant screening of loop A, single-stranded stretch, and the terminator loop. Labeled LhrC4 and the mutant derivatives were tested for their ability to interact with unlabeled lmo0484 RNA. The mutated regions are shown in red in the sRNA sketches. LhrC4: wild-type LhrC4; LhrC4_mut_2: mutation in single-stranded stretch; LhrC4_mut_3: mutation in terminator loop; LhrC4_mut_4: mutation in loop A. The fraction of unbound LhrC4 is shown below each lane. (B) Predicted basepairing between the SD region of lmo0484 mRNA and loop A of LhrC4. The mutated nucleotides are shown in bold and the sequences of the minimal mutant variants lmo0484_MUT and LhrC4_loopA_MUT are indicated. (C) Labeled lmo0484 RNA and lmo0484_MUT were each incubated with increasing concentrations of unlabeled LhrC4 or the mutant variant LhrC4_loopA_MUT. The fraction of unbound lmo0484 RNA is shown below each lane.