Zhu Qin-Long, Yang Zhong-Fang, Zhang Qun-Yu, Chen Le-Tian, Liu Yao-Guang
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
Gene. 2014 Sep 10;548(1):39-42. doi: 10.1016/j.gene.2014.07.004. Epub 2014 Jul 8.
A robust strategy for multi-type plasmid modifications is developed based on the isothermal in vitro recombination technology, by which any combination of the sequence modifications can be efficiently achieved in plasmids at any desired position in a seamless manner. As an example, we showed that a plasmid modification with insertion of a GFP gene, deletion of a 623-bp fragment, and substitution of an ampicillin resistance gene by a kanamycin resistance gene was accomplished simultaneously by this method. Therefore, the isothermal in vitro recombination-based multi-type plasmid modification strategy is a useful approach for broad application prospects in molecular biology studies.
基于等温体外重组技术开发了一种用于多类型质粒修饰的强大策略，通过该技术可以无缝地在质粒的任何所需位置高效实现序列修饰的任何组合。例如，我们表明通过该方法可以同时完成插入GFP基因、缺失623 bp片段以及用卡那霉素抗性基因替换氨苄青霉素抗性基因的质粒修饰。因此，基于等温体外重组的多类型质粒修饰策略是一种在分子生物学研究中具有广阔应用前景的有用方法。
