1.遵义医科大学珠海校区,珠海 519040;2.珠海市中药基础与应用研究重点实验室,珠海 519040
国家自然科学基金(31560244),贵州省科技合作计划项目(黔科合LH字[2015]7547),遵义市“15851人才精英工程”科研项目和珠海市中药基础与应用研究重点实验室开放课题(ZYKL-2016K2)资助项目.
1.Zhuhai Campus of Zunyi Medical University, Zhuhai 519040, China;2.Zhuhai Key Laboratory of Fundamental and Applied Research in Traditional Chinese Medicine, Zhuhai Campus of Zunyi Medical University, Zhuhai 519040, China
This work was supported by grants from The National Natural Science Foundation of China(31560244), Science and Technology Cooperation Project of Guizhou Province(LH20157547), “15851 Talent Elite Project” of Zunyi City, Opening Project of Zhuhai Key Laboratory of Fundamental and Applied Research in Traditional Chinese Medicine(ZYKL-2016K2).
无限制克隆(restriction-free cloning,RFC)是近年来建立起来的一种简单通用的DNA克隆技术,它可以精确地将目的片段插入到质粒内任意位置,是一种不受酶切位点、连接酶效率、目的基因长度或载体序列等条件限制的新型DNA重组技术.与其他多种克隆方法相比,RFC技术拥有不可替代的优势.本文在总结国内外RFC技术研究的基础上,系统阐述了RFC技术的原理、特点和在克隆方面的研究进展,并探讨其在分子生物学和合成生物学等领域的应用价值.
Restriction free cloning(RFC) is a simple and universal DNA cloning technology developed in recent years, which can accurately insert the target DNA sequence into any position of any plasmids. It is a novel DNA recombination method that is not limited by restriction endonuclease, efficiency of ligase, length of target DNA sequence or vector sequence. Compared with other cloning methods, RFC technology has irreplaceable advantages. On the basis of summarizing the researches of RFC at home and abroad, this manuscript systematically introduces the action principle, characteristics and research progress of RFC technology and discuss its application value in molecular biology and synthetic biology.
杨传辉,卢荣锐,杨愈丰.无限制克隆技术研究进展及其应用[J].生物化学与生物物理进展,2019,46(7):719-726
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