【摘 要】
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Nicking endonucleases (NEases) are sequence-specific and strand-specific nicking enzymes that hydrolyze only one strand of ds-DNA targets.Natural NEases Nb.
【机 构】
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NewEnglandBiolabs,Inc.240CountyRoad,Ipswich,MA01938
【出 处】
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第五届中国工业生物技术发展高峰论坛
论文部分内容阅读
Nicking endonucleases (NEases) are sequence-specific and strand-specific nicking enzymes that hydrolyze only one strand of ds-DNA targets.Natural NEases Nb.BsrDI, Nb.BtsI, Nt.CviPII, and Nt.CviQII were cloned, expressed, and purified.Other engineered NEases are Nb.Bsml, Nt.BsrDl, Nt.BtsI, Nt.BsmAl, and Nt.BspQI that are useful for DNA nicking/labeling, and for single molecule studies.Nt.CviPII and Nt.BspQI have been used for strand-displacement amplification in conjunction with a DNA polymerase.We also constructed chimeric endonucleases by fusion of BmrI cleavage domain to other DNA binding proteins such as controller protein (C protein), catalytic-deficient NotI variant, and 5mC binding domain of MeCP2.The chimeric endonucleases can be targeted to C protein binding box, outside of Notl sequence, and near 5mC sites.
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