RedET重组及其在生物医学中的应用解析.docVIP

21卷3期2005年5月 生　物　工　程　学　报 Chinese Journal o f Biotechnology V ol. 21　N o. 3 May 　2005 R ed ΠET 重组及其在生物医学中的应用 R ed ΠET R ecombination and its Biomedical 王军平 13 　 , 张友明 2 W ANGJun 2Ping 13and 　ZH ANG Y ou 2Ming 2 11, 重庆因桥研究室, 11State K ey Laboratory o f , and Injury , Institute o f Combined Injury , College o f Preventive Medicine , The Third Military Medical Univer sity , Chongqing 　400038, 21G ene Bridge G mbH , Dresden 01307, G ermany αβ系统而建立的DNA 工程平台—摘　要　通过应用Rac 噬菌体的RecE ΠRecT 和λ噬菌体的Red ΠRed ——Red ΠET 重组, 是一种不依赖于限制性内切酶的分子克隆新技术。运用该技术能够介导PCR 产物或寡核苷酸对目标基因进行剪切、插入、融合及突变等多种操作, 在生物医学领域里具有广阔的应用前景, 尤其在基因组功能研究中对BACs 、PACs 和细菌染色体的打靶修饰以及基因敲除动物DNA 靶分子的快速构建等方面最有效。随着Red ΠET 重组的推广与应用, 该技术本身也在不断被改进, 在工作效率得到显著提高的同时, 其操作也变得更加简单、省时、省力。结合自身的一些研究结果, 对Red ΠET 重组的技术特点、发展现状和在生物医学中的应用进行了详细阐述。关键词　Red ΠET 重组, DNA 修饰, BACs , 噬菌体 中图分类号　Q78　　文献标识码　A 　　文章编号100023061(2005 0320502205 Abstract 　Red ΠET recombination , a powerful hom olog ous recombination system based on the Red operon of λphage or RecE ΠRecT from Rac phage , provides an innovative approach for DNA engineering. Deletion , insertion and mutation can be quickly and precisely performed on the target gene mediated by Red ΠET recombination with PCR derived DNA fragments or olig onucleoti 2des. This technical platform has extensive applications in biomedical field including bacterial artificial chrom osome m odification , gene knock 2out construction and genetic m odification of E . coli strains as well as some other kinds of m icroorganisms. Recently , Red ΠET recombination was im proved in several aspects so that it becomes m ore powerful and maneuverable. The characteristic and development of Red ΠET recombination and its biomedical applications were described in this review. K ey w ords 　Red ΠET recombination , DNA m odification , bacterial artificial chrom osomes , phage 　　随着包括人类基因组计划在内的各种基因组测序工程的实施与完成, 以序列信息为基础的基因组功能研究随即成为生命科学领域的又一重大课题[1]。相对于测序工程, 基因组的功能研究就更加复杂和困难。对于某一特定基因, 要想彻底了解其生物学作用, 往往需要对包含基因完整信息的基 因组DNA 进行克隆、删除、突变等多种