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. 2013 May 23;8(5):e64609. doi: 10.1371/journal.pone.0064609

Figure 3. Electrophoretic mobility shift assays using CrcŹ RNA.

Figure 3

Electrophoretic mobility shift assay of 10 nM 5′-end labeled CrcZ′ RNA with increasing amounts of His-Crc purified from the E. coli strain Rosetta™ (DE3)(pLysS pETM14lic-His6Crc) by one-step NAC (A) and by one-step NAC followed by SEC (B), respectively. EMSA assay employing the His-Crc protein purified from the P. aeruginosa strain PAO1(pME9670) by one-step NAC (C), the protein eluate obtained after one-step NAC from strain Rosetta™ (DE3)(pLysS, pETM14lic) (mock; no Crc protein) (D) and the His-Crc protein from the E. coli hfq- strain JW4130(pME9670) by one-step NAC (E). Lane 1, no protein was added to labeled CrcZ` RNA. Lanes 2-4, the protein fractions were added in 50, 100 and 200-fold molar excess over labeled RNA. In the case of the mock preparation (D), the same amount of protein was added to RNA as in the experiments shown in panels A, B and C.