工业微生物代谢途径调控的基因敲除策略

Gene knockout strategies for metabolic pathway regulation in industrial microbes

摘要:

基因敲除技术是一项重要的分子生物学技术，在工业微生物代谢工程中具有广泛应用。以下从基因敲除技术的遗传重组原理出发，总结了基因敲除策略的类型、特征和应用，重点介绍了采用线性双链DNA的λ Red同源重组系统、使用环状质粒载体介导的单交换或双交换同源重组策略以及采用转座酶介导的转座重组等几种主要的基因敲除方法，进一步展望了基因敲除技术的发展前沿和应用前景。

Abstract:

Gene knockout, an important technology in molecular biology, has been broadly applied in industrial microbial metabolic engineering. From the basic mechanism of DNA recombination, we summarized and compared in this review different gene knockout strategies. Three most hot and important approaches, including the λ Red recombination system using the linear dsDNA as recombination substrate, the single or double crossover homologous recombination using the circular plasmid DNA as substrate, and the transposase mediated transposition recombination, were summarized in detail. Developing frontiers and application prospects of gene knockout were further discussed.