| 论文题目: | An efficient blue-white screening based gene inactivation system for Streptomyces |
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| 作者: | Li Pengwei Li Jine, Guo Zhengyan, Tang Wei, Han Jianshan, Meng Xiangxi, Hao Tingting, Zhu Yaxin, Zhang Lixin, Chen Yihua*. |
| 联系作者: | |
| 刊物名称: | Appl Microbiol Biotechnol |
| 期: | 99 |
| 卷: | 4 |
| 页: | 1923-33 |
| 年份: | 2015 |
| 影响因子: | 4.138 |
| 论文下载: | 下载地址 |
| 摘要: | Streptomyces is studied intensively for its outstanding ability to produce bioactive secondary metabolites and for its complicated morphological differentiation process. A classical genetic manipulation system for Streptomyces has been developed and widely used in the community for a long time, using antibiotic resistance markers to select for double-crossover mutants. The screening process is always laborious and time-consuming. However, the lack of a suitable chromogenic reporter for Streptomyces has limited the use of color-based screening system to simplify the selection process for double-crossover mutants. In this study, a blue reporter system for Streptomyces has been established by mining an indigoidine synthetase gene (idgS) from Streptomyces lavendulae CGMCC 4.1386, leading to the development of a time-saving gene inactivation system for Streptomyces by simple blue-white screening. A series of Streptomyces suicide and temperature-sensitive plasmids containing the idgS reporter cassette were constructed and used successfully to inactivate genes in Streptomyces, allowing a simple and efficient screening method to differentiate the colonies for double-crossover (white) and single-crossover (blue) mutants. Inactivation of the putative gamma-butyrolactone synthase gene afsA-y via the idgS-based blue-white screening method revealed that the paulomycin production is negatively controlled by afsA-y in Streptomyces sp. YN86. |
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