State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.
University of Chinese Academy of Sciences, Beijing, 100049, China.
Sci China Life Sci. 2022 Jun;65(6):1248-1256. doi: 10.1007/s11427-021-1998-8. Epub 2021 Oct 14.
Amplification of biosynthetic gene clusters is important to increase secondary metabolite production. However, the copy number of amplified gene clusters is difficult to control precisely. In this study, the tandem amplification of a 70 kb bleomycin biosynthetic gene cluster was precisely regulated through the combined strategy of a ZouA-dependent DNA amplification system and double-reporter-guided recombinant selection in Streptomyces verticillus ATCC15003. The production of bleomycin in the recombinant strain containing six copies of the bleomycin gene cluster was 9.59-fold higher than that in the wild-type strain. The combined strategy used in this study is powerful and applicable for precisely regulating the amplification of gene clusters and improving the corresponding secondary metabolite production.
生物合成基因簇的扩增对于增加次生代谢产物的产量很重要。然而，扩增基因簇的拷贝数很难精确控制。在这项研究中，通过结合 ZouA 依赖的 DNA 扩增系统和双报告基因指导的重组选择策略，精确调控了委内瑞拉链霉菌 ATCC15003 中 70kb 博莱霉素生物合成基因簇的串联扩增。含有六个博莱霉素基因簇的重组菌株的博莱霉素产量比野生型菌株提高了 9.59 倍。本研究中使用的组合策略功能强大，适用于精确调控基因簇的扩增，提高相应的次生代谢产物产量。
