Red/ET同源重组技术及其在微生物基因组挖掘中的应用进展

Advances in Red/ET recombineering and its application for microbial genome mining

摘要:

Red/ET同源重组技术（Red/ET recombineering）是由来源于大肠杆菌λ噬菌体的蛋白对Redα/Redβ或来源于Rac原噬菌体的蛋白对RecE/RecT所介导的基于短同源臂（40-50 bp）的同源重组技术，能对宿主DNA序列进行快速、高效、精确的修饰和操作。本文主要综述了2010年以来Red/ET同源重组技术在大肠杆菌及其他细菌中的研究进展，同时简要介绍了该技术在微生物基因组挖掘，尤其是在微生物基因簇的异源表达领域的应用进展。

关键词:Red/ET同源重组技术;直接克隆;遗传工程;基因组挖掘;次级代谢产物;异源表达

Abstract:

Red/ET recombineering is an in vivo homologous recombination-based genetic engineering method used primarily in Escherichia coli by using short homology arms (40-50 bp), based on the expression of either redα/redβ from the Red operon of λ phage or the analogous recE/recT from Rac prophage that are located in the E. coli chromosome. It can rapidly, efficiently and accurately modify and manipulate genomic and episome DNA. In this review, we introduce the progress of Red/ET recombineering in E. coli and other bacteria, and its application in microbial genome mining, especially in the field of heterologous expression of microbial biosynthetic gene clusters.

Key words:Red/ET recombineering;direct cloning;genetic engineering;genome mining;secondary metabolites;heterologous expression