Li Xuegang, Jin Jiacheng, Guo Zhanyong, Liu Liangwei
The Life Science College, Henan Agricultural University, Zhengzhou 450002, China.
National Key Laboratory of Wheat and Maize Crop Science, Henan Agricultural University, Zhengzhou 450002, China.
Int J Biol Macromol. 2022 Jun 1;209(Pt A):1319-1326. doi: 10.1016/j.ijbiomac.2022.04.094. Epub 2022 Apr 20.
Developing for almost half a century, plasmid-construction has explored more than 37 methods. Some methods have evolved into new versions. From a global and evolutionary viewpoint, a review will make a clear understand and an easy practice for plasmid-construction. The 37 methods employ three principles as creating single-strand overhang, recombining homology arms, or serving amplified insert as mega-primer, and are classified into three groups as single strand overhang cloning, homologous recombination cloning, and mega-primer cloning. The methods evolve along a route for easy, efficient, or/and seamless cloning. Mechanism of plasmid-construction is primer annealing or/and primer invasion. Scar junction is a must-be faced scientific problem in plasmid-construction.
经过近半个世纪的发展，质粒构建已探索出37种以上的方法。一些方法已演变成新版本。从全局和进化的角度来看，一篇综述将有助于清晰理解和轻松实践质粒构建。这37种方法采用了三种原理，即创建单链突出端、重组同源臂或使用扩增的插入片段作为巨型引物，并分为三组，即单链突出端克隆、同源重组克隆和巨型引物克隆。这些方法沿着一条实现简便、高效或/和无缝克隆的路线发展。质粒构建的机制是引物退火或/和引物侵入。疤痕连接是质粒构建中必须面对的科学问题。
