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OFX-mediated DNA cleavage assay. Supercoiled pBR322 (0.3 μg) was incubated with GyrA (50 ng) and GyrB (50 ng) in the presence of the indicated concentration of OFX. DNA cleavage assay was performed with combinations consisting of WT GyrB-WT GyrA (A), GyrA-Ala91Val (B), GyrA-Asp95Gly (C), and GyrA-Asp95Asn (D). R, L, and SC, relaxed, linear, and supercoiled pBR322 DNA, respectively.
