Figure 5.

Comparison of DNA cleavage by FokI and FokD483A, R487A. The cleavage reactions were performed for 1 h at 37°C in 30 μl of 20 mM Tris-acetate buffer (pH 7.9), 10 mM magnesium acetate, 50 mM potassium acetate, 1 mM DTT, and 100 μg/ml BSA, containing 20 nM of (A and C) supercoiled plasmid pSKFok1 or (B and D) supercoiled plasmid pSKFok0 and either (A and B) FokI or (C and D) FokD483A, R487A. The concentrations of FokI and FokD483A, R487A are indicated. (CC), covalently closed form of plasmid DNA; (OC), open circle form; (L), linear form.