Red/ET重组及其在生物医学中的应用

Red/ET Recombination and its Biomedical Applications

通过应用Rac噬菌体的RecE RecT和λ噬菌体的RedαRedβ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、省时、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。

Red/ET recombination, a powerful homologous recombination system based on the Red operon of λ 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 oligonucleotides. 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.