微生物基因打靶系统的研究进展  

Advances in gene targeting system of microbe

作　　者：谭晓红[1] 程萱[1] 杨晓军[1] 

机构地区：[1]事医学科学院生物工程研究所,北京100071

出　　处：《生物技术通讯》2004年第3期263-266,共4页Letters in Biotechnology

摘　　要：基因打靶技术是微生物功能基因组学研究的有力工具之一,通过定向改变微生物的遗传信息可以对目的基因进行有效的功能分析。在大肠杆菌中研究较多的是转座子突变系统、RecBCD-sbcB重组系统、RecA依赖的重组打靶系统、Chi位点刺激的重组、利用单链DNA进行的重组工程。在酵母中进行基因打靶的策略主要是转座子标记的突变、基于PCR方法的基因删除和转化相关重组。在其他微生物中主要应用转座子突变和自杀载体进行基因打靶。近年来,噬菌体重组系统的发展更使对微生物基因打靶系统的研究进入了新的阶段,主要包括Rac编码的RecET系统、Red重组系统和噬菌体退火蛋白介导的单链寡核苷酸重组系统。Gene targeting,an approach that has been used to modify the genetic information of organism in a predeter-mined way,is a powerful tool for studying the functional genomics of microbe.In E.coli,investigation is focused on sys-tems of transposon mutagenesis,RecBCD-sbcB-dependent recombination,RecA-dependent recombination,Chi-stimulated recombination,ssDNA recombination.The primary strategies of gene targeting in yeast are transposon tagging and mutagen-esis,genome-wide gene deletions and transformation-associated recombination.Transposon mutagenesis and suicide vectors are mainly used for gene targeting in other microbes.The recently developed phage-based homologous recombination sys-tems have opened new avenues for gene targeting in microbes,including rac-encoded RecET system,Red recombination and oligonucleotide-directed mutagenesis promoted by phage annealing proteins.

关 键 词：微生物 基因打靶 功能基因组学 同源重组 转座子 

分 类 号：Q78[生物学—分子生物学]