文献类型： 外文期刊
作者： Yan, Pu 1 ; Gao, XinZheng 3 ; Shen, Wentao 1 ; Zhou, Peng 1 ; Duan, Jun 2 ;
作者机构： 1.Chinese Acad Trop Agr Sci, Inst Trop Biosci & Biotechnol, Haikou 571101, Peoples R China
2.Chinese Acad Sci, S China Bot Garden, Guangzhou 510650, Guangdong, Peoples R China
3.Hainan Med Coll, Coll Sci, Haikou 571101, Peoples R China
关键词： Golden Gate cloning;Multiple mutagenesis;Parallel assembly;Plasmid
期刊名称：ANALYTICAL BIOCHEMISTRY （ 影响因子：3.365； 五年影响因子：3.049 ）
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收录情况： SCI
摘要： A parallel assembly method for multiple site-directed mutagenesis of plasmids was developed here based on Golden Gate cloning. It takes advantage of type IIs restriction enzymes and T4 DNA ligase to assemble multiple DNA fragments into a plasmid by a defined order. This method can accommodate multiple plasmid mutagenesis at any desired position with all three sequence modification types (substitution, deletion, and insertion) simultaneously. Furthermore, it can be used to create otherwise difficult-to-make mutants - larger deletions and insertions and mutagenesis on larger plasmids. The processes of mutagenesis can be completed quickly by a single restriction-ligation reaction.
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