蛋白质体外进化建库技术研究

Studies on Constructing Libraries of Directed Evolution of Proteins in vitro

蛋白质体外进化技术是蛋白质工程发展的一个里程碑,也是改造蛋白质的一种有效工具。它不仅具有重要的应用价值,而且有助于蛋白质结构与功能的研究。通过蛋白质体外进化技术已成功地改造了许多蛋白质,有些已应用于工农业生产。体外进化技术分为两步:建库和筛选。本文主要对蛋白质体外进化策略及对体外随机突变技术、DNA重组技术、利用活细胞自身修复系统构建突变文库等几种定向进化突变文库建立技术进行了介绍与论述,同时还对蛋白质体外进化技术的应用及与其它学科结合的研究前景进行了分析,为获得具有改进功能或全新功能的蛋白质提供理论基础。

Directed evolution of proteins in vitro is a milestone achievement in protein engeering,which is an effective tool for protein modification. It is of wide application,and is regarded quite helpful for studying protein sturcture and function. Up to now,many proteins have been successfully modified based on this technique,some of which have already been applied in industrial and agricultural production. The technique consists of two steps：libiary construction and screening. In this review,we mainly introduce and review the strategies of protein evolution and sum up several directed evolution techniques for generating mutation libraries,such as random mutagenesis in vitro technique,recombinant DNA and self-repair system of living cell. At the same time,we analyze the prospect regarding the application and interdisciplinary research of this technique. This review would offer theoretical foundation for the development of proteins with improved or bran-new function.