基于PCR的体外诱变技术

PCR-based site-directed mutagenesis

定点突变技术是蛋白质工程中应用的重要技术之一,可以有目的改变DNA序列中的碱基。它不仅可以阐明基因表达的调控机理,还可用研究蛋白质结构与功能间的关系,改造天然蛋白质,使之更符合应用需求。自从PCR技术发明以来,便被用于进行定点突变。综述了各种基于PCR技术的定点突变方法,并与其他定点突变方法进行对比。

In vitro site-directed mutagenesis is an invaluable technique for studying protein structure-function relationships and gene expression,Designing new proteins, and for Identifing enzyme active sites. Oligonucleotide-directed site-specific mutagenesis is previously used routinely to introduce desired mutations into target DNA sequences.PCR-based methods are currently widely used because of fheir flexibility and the yield of mutants approaches 100%.Recent advances and applications of PCR-based site-directed mutagenesis are reviewed and discussed in the article.