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(A) DNA nicking or restriction activity of BtsI A/B mutants. B cat* = catalytic-deficient B subunit. ϕX174 RF DNA (1 BtsI site) was used as the substrate for nicking or restriction assay. NC, nicked-circular DNA; L, linear DNA; SC, supercoiled DNA. (B) Run-off sequencing of gel-purified, nicked-circular DNA. The expected nicking specificity of Nt.BtsI is GCAGTGNN↓ (BtsI site and the adjacent sequence are shown at the bottom of the figure).
