基于抗性恢复的高效克隆超短片段重组质粒的构建  

Cloning short gene fragment and constructing recombinant plasmid based on restoration of antibiotic resistance

作　　者：刘美丽[1] 罗砚曦[1] 赵铮蓉[1] 阎辉[1] 

机构地区：[1]浙江省医学科学院药物研究所,浙江杭州310013

出　　处：《生物工程学报》2017年第7期1190-1197,共8页Chinese Journal of Biotechnology

基　　金：浙江省医药卫生科研项目(No.2014KYA041)资助~~

摘　　要：分子克隆作为一种常规技术被广泛应用于DNA及蛋白质的研究。在传统的分子克隆中,主要通过限制性内切酶先分别消化目的 DNA片段及载体,再纯化回收,然后用DNA连接酶将二者连接。而对一些超短基因片段(<300 bp),通过酶切及切胶纯化后,回收率极低,导致插入表达载体比较困难。文中介绍了一种新的利用质粒抗性恢复进行克隆的方法,大大提高了克隆效率,为短基因片段的分子克隆提供了一种高效的方法。Molecular cloning is one of the most important and widely used technologies in molecular biology research. Generally, the target DNA fragment and the vector are separately digested by restriction enzyme, then purified and recovered, and then ligated with DNA-ligase. For some very short gene fragments （〈300 bp）, the recovery efficiency of the purified fragment is very low after digestion and cleavage, leading to the difficulty in its inserting into the expression vector. To address this issue, we developed a cloning method based on restoration of antibiotic resistance in constructing recombinant plasmid, which proved highly efficient in cloning very short gene fragments.

关 键 词：分子克隆 构建 短基因片段 

分 类 号：Q78[生物学—分子生物学]