文献类型： 外文期刊
第一作者： Yan, Pu
作者： Yan, Pu;Zeng, Yanjing;Shen, Wentao;Tuo, Decai;Li, Xiaoying;Zhou, Peng
作者机构：
关键词： molecular cloning; standardized cloning; Gibson assembly-derived cloning; vector construction; DNA assembly; prokaryotic expression; plant gene expression
期刊名称：FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY （ 影响因子：5.89； 五年影响因子：5.973 ）
ISSN： 2296-4185
年卷期： 2020 年 7 卷
页码：
收录情况： SCI
摘要： Molecular cloning is one of the most fundamental technologies in molecular biology, and has been critical for driving biotechnological advances. In this study, we have developed a novel method for standardized molecular cloning. The cloning technique known as "Nimble Cloning" uses the restriction enzyme, SfiI, in combination with the T5 exonuclease, to linearize the vector and generate 3 '-overhangs simultaneously. Both PCR products and plasmids can be used for the cloning reaction in the Nimble Cloning system. The cloning system is highly efficient, suitable for gene expression in both prokaryotic and eukaryotic expression systems, and enables the reuse of DNA fragments or plasmid entry clones. Nimble Cloning is applicable for the cloning of single or multiple fragments, as well as multi-site cloning. Due also to its simplicity and versatility, the cloning method has great potential for the modular assembly of DNA constructs.
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