Article
Increasing cleavage specificity and activity of restriction endonuclease KpnI.
Nucleic acids research - 1 Nov 2013
Vasu Kommireddy, Nagamalleswari Easa, Zahran Mai, Imhof Petra, Xu Shuang-Yong, Zhu Zhenyu, Chan Siu-Hong, Nagaraja Valakunja
Abstract excerpt
Restriction enzyme KpnI is a HNH superfamily endonuclease requiring divalent metal ions for DNA cleavage but not for binding. The active site of KpnI can accommodate metal ions of different atomic radii for DNA cleavage. Although Mg(2+) ion higher than 500 μM mediates promiscuous activity, Ca(2+) suppresses the promiscuity and induces high cleavage fidelity. Here, we report that a conservative mutation of the...
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