不依赖连接反应的高通量克隆方法  

Methods for High-Throughput Cloning without Ligation Reaction

作　　者：李华琴[1] 林陈水[1] 张文倩[1] 

机构地区：[1]浙江工业大学药学院,浙江杭州310014

出　　处：《氨基酸和生物资源》2013年第4期43-46,共4页Amino Acids & Biotic Resources

摘　　要：基因表达及功能分析需要高通量的表达系统,因而简单、经济且高效地将PCR产物或其他来源的目的基因片段构建到质粒载体中就成为了亟需解决的问题。传统克隆方法使用限制性内切酶和连接酶,其缺点是受到基因序列中原有的酶切位点的限制以及依赖连接酶导致的较低的连接转化率。为了克服这些缺点,一些不需要限制酶和连接酶的克隆方法被开发出来并运用到基因克隆中,获得了更佳的实验结果。该文就LIC、UDG克隆和Gateway重组克隆这三种不依赖连接反应的高通量克隆方法的原理及特性进行介绍。With the development of gene cloning and expression,how to put PCR products or other source of purpose gene fragment into the plasmid vector simply,economically and efficiently is a problem for people to solve.The disadvantages of traditional practices using restriction enzymes and ligase are low translating connection by limit of restrict enzyme sites and use of ligase.In order to overcome these shortcomings,many experimenters tried to use some cloning methods without restriction enzyme and ligase in gene clonging,to obtain better results.This paper intoduces the principle and characteristics of three methods for high-throughput cloning,which contain ligation independent cloning,gateway recombinational cloning and uracil DNA glycosylase cloning.

关 键 词：不依赖连接克隆 Gateway重组克隆 UDG克隆 

分 类 号：Q784[生物学—分子生物学]