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. 1998 Sep 1;95(18):10570–10575. doi: 10.1073/pnas.95.18.10570

Figure 4.

Figure 4

Stimulation of FokI endonuclease by (A) FokN13Y, (B) FokCD, or (C) FokCD/D450A. Reactions were performed at 37°C in 70 μl of 20 mM Tris-acetate buffer (pH 7.9), 10 mM magnesium acetate, 50 mM potassium acetate, 1 mM DTT, 100 μg/ml BSA, and 6.3 nM linearized pSKFok1 DNA; 10-μl samples were withdrawn at the indicated times from the reactions that contained: (A) either 500 nM FokN13Y, 1.0 nM FokI, or 1.0 nM FokI + 500 nM FokN13Y; (B) either 500 nM FokCD, 0.5 nM FokI, or 0.5 nM FokI + 500 nM FokCD; and (C) either 500 nM FokCD/D450A, 1.0 nM FokI, or 1.0 nM FokI + 500 nM FokCD/D450A. Samples were quenched by adding 5 μl of stop solution and were analyzed by electrophoresis on 1% agarose gel.