SinR’ interacts with SinR.

<p><b>(A)</b><i>In vitro</i>, protein-protein interactions indicate that SinR’ binds tightly to SinR. GST-tagged SinR’ protein was incubated with SinR-6His proteins and purified using Ni++ agarose affinity columns. The elutes were probed with anti-GST and with anti-His antibodies. Lanes details are as follows: Input 1: Mixture of SinR’-GST expressing <i>E</i>.<i>coli</i> lysate with purified SinR-6His. 1.Input 2: Mixture of GST expressing <i>E</i>. <i>coli</i> lysate with purified SinR-6His. 2. Unbound from input 1 after passing through Ni⁺⁺ column. 3. Unbound from input 2 after passing through Ni⁺⁺ column. 4. Elute with 50 mm imidazole (SinR’-GST + SinR-6His). 5. Elute with 50 mM imidazole (GST+SinR-6His) 6. Elute with 200 mM imidazole (SinR’-GST + SinR-6His). 7. Elute with 200 mM imidazole (GST+SinR-6His). * indicates SinR-His dimer. <b>(B)</b> Interactions of SinR with <i>codY</i> promoter region. EMSA analysis of SinR-6His, SinR’-6His, a mixture of SinR’-6His and SinR-His binding to <i>codY</i> probe.