Structure-guided design and rapid cell-free screening of a superior restriction endonuclease DpnI mutant to enhance the specific activity and efficiency of site-directed mutagenesis.

Abstract excerpt

Restriction endonuclease DpnI is a critical tool for eliminating parental templates in site-directed mutagenesis (SDM); however, the wild-type enzyme exhibits low efficiency at low substrate concentrations. In this study, we introduced a lysine mutation into the C-terminal winged helix (wH) domain of DpnI via structure-guided design to enhance electrostatic interactions and established a rapid screening platform...

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