Red／ET重组及其在生物医学中的应用.pdfVIP

2l卷 3期 生 物 工 程 学 报 Vo1．2l No．3 2005年5月 Chinese Journal of Biotechnology Mav 2005 Red／ET重组及其在生物医学中的应用 Red／ET Recombination and its Biomedical Applications 王军平 ，张友明 WANG Jun—Ping and ZHANG You—Ming 1．第三军医大学预防医学系全军复合伤研究所，创伤烧伤复合伤国家重点实验室，重庆 400038 2．基因桥研究室，德累斯顿 01307，德国 1．State Key Laboratory ofTrauma，Burns and Combined l埔u ，Institute ofCombined l畸“ry，College ofPreventive Medicine，The Third Military Medical University，Chongqing 400038，China 2．Gene Bridge GmbH，Dresden 01307，Germany 摘 要 通过应用Rac噬茵体的RecE／RecT和 噬茵体的Reda／Red~3系统而建立的DNA工程平台——Red，ET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／ET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、唐时、省力。结合自身的一些研究结果，对Red／ET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／ET重组，DNA修饰，BACs，噬茵体 中图分类号 Q78 文献标识码 A 文章编号1000—306l【2005)03—0502—05 Abstract Red／ET recombination．a powerful homologous recombination system based on the Red Ol~ron of phage or RecE／ RecT 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／ET recombination with PCR derived DNA fragments or oligonucleoti— des．This technical platform has extensive applications in biomedical field including bacterial artificial chromosome modification， gene knock—out construction and genetic modification of E．coli strains as well as some other kinds of microorganisms．Recently， Red／ET recombination Was improved in several aspects SO that it becomes more powerful and maneuverable．The characteristic and development of Red／ET recombination and its biomedical applications were described in this review Key words Red／ET recombination，DNA modification，bacterial artificial chromosomes，phage 随着包括人类