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. Author manuscript; available in PMC: 2012 Jan 7.
Published in final edited form as: J Mol Biol. 2010 Oct 26;405(1):185–200. doi: 10.1016/j.jmb.2010.10.029

Figure 4.

Figure 4

Competitive DNA cleavage assay by wild-type I-SceI and mutant derivatives. A) Two linearized plasmid substrates containing either the wild-type I-SceI recognition sequence (A/T+4) or the C/G+4-substituted site (10 nM) were incubated for 20 min at 25° C with different concentrations of wild-type I-SceI (0, 5 nM, 10 nM, 20 nM, 40 nM, 80 nM, 160 nM, 320 nM, or 640 nM). The uncleaved DNA substrates were separated from the cleavage products by agarose gel electrophoresis, the gels were stained with ethidium bromide and the amounts of each band were quantitated from digital images. B) As in panel A) using the G100T I-SceI derivative. C) As in panel A) using the K86R/G100T protein. D) Average C50 values from duplicate DNA cleavage experiments.