蛋白质定向进化的高通量筛选方法

定向进化是改造蛋白质分子的有效新策略。创建突变库的方法已经有很多而且比较通用,而建立有效的高通量的筛选方法是蛋白质定向进化成功的关键。本文综述了近几年发展起来的用于定向进化的高通量的文库筛选方法,介绍了各种展示技术及流式细胞分选技术的原理及其在文库筛选中的应用,分析了存在的问题及发展趋势。

HIGH- THROUGHPUT SCREENING METHODS IN THE DIRECTED EVOLUTION OF PROTEINS

Directed evolution is a new powerful strategy for engineering proteins.Many feasible methods for generating gene libraries are developed.Therefore developing the high-throughput screening methods for desired mutants is the key to the success in directed evolution.In this article,we summarize the high-throughput screening methods developed resent years such as FACS(Fluorescence-activated cell sorting) and the display technology including phage display,cell-surface display,ribosome display and mRNA-peptide fusion.Their principles and application in screening for desired mutants from mutant libraries are introduced.We discuss the unresolved questions and promising prospect in the screening methods.