基于 CRISPR/Cas9 的方法用于失活放线菌生物合成基因和阐明功能。
CRISPR/Cas9-Based Methods for Inactivating Actinobacterial Biosynthetic Genes and Elucidating Function.
机构信息
Biophore - DMF, University of Lausanne, Lausanne, Switzerland.
School of Life Sciences, University of Warwick, Coventry, UK.
出版信息
Methods Mol Biol. 2022;2489:201-222. doi: 10.1007/978-1-0716-2273-5_11.
The CRISPR/Cas9 technology allows fast and marker-less genome engineering that can be employed to study secondary metabolism in actinobacteria. Here, we report a standard experimental protocol for the deletion of a biosynthetic gene in a Streptomyces species, using the vector pCRISPomyces-2 developed by Huimin Zhao and collaborators. We also describe how carrying out metabolite analysis can reveal the putative biosynthetic function of the inactivated gene.
CRISPR/Cas9 技术可实现快速、无标记的基因组工程，可用于研究放线菌中的次生代谢。本研究报告了使用胡敏教授及其合作者开发的 pCRISPomyces-2 载体对链霉菌属物种中的生物合成基因进行缺失的标准实验方案。我们还描述了如何进行代谢产物分析以揭示失活基因的潜在生物合成功能。
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