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Recombineering for Genetic Engineering of Natural Product Biosynthetic Pathways.
Trends in Biotechnology ( IF 14.3 ) Pub Date : 2020-01-20 , DOI: 10.1016/j.tibtech.2019.12.018
Muhammad Nazeer Abbasi 1 , Jun Fu 1 , Xiaoying Bian 1 , Hailong Wang 1 , Youming Zhang 1 , Aiying Li 1
Affiliation  

Microbial genomes encode many cryptic and uncharacterized biosynthetic gene clusters (BGCs). Exploiting this unexplored genetic wealth to discover microbial novel natural products (NPs) remains a challenging issue. We review homologous recombination (HR)-based recombineering, mediated by the recombinases RecE/RecT from Rac prophage and Redα/Redβ from lambda phage, which has developed into a highly inclusive tool for direct cloning of large DNA up to 100 kb, seamless mutation, multifragment assembly, and heterologous expression of microbial NP BGCs. Its utilization in the refactoring, engineering, and functional expression of long BGCs for NP biosynthesis makes it easy to elucidate NP-producing potential in microbes. This review also highlights various applications of recombineering in NP-derived drug discovery.



中文翻译:

天然产物生物合成途径的基因工程重组。

微生物基因组编码许多秘密的和未表征的生物合成基因簇(BGC)。利用这种未开发的遗传财富来发现微生物新颖的天然产物(NPs)仍然是一个具有挑战性的问题。我们综述了基于同源重组(HR)的重组,该重组是由来自Rac prohage的RecE / RecT和来自λ噬菌体的Redα/Redβ介导的,已发展成为一种高度包容的工具,可直接克隆高达100 kb的大DNA,进行无缝突变,NP的多片段组装和异源表达。它在长BGC用于NP生物合成的重构,工程和功能表达中的利用,使其易于阐明微生物中产生NP的潜力。这篇综述还重点介绍了重组技术在NP衍生药物发现中的各种应用。

更新日期:2020-01-20
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