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通过可逆同源重组开关,促进稳定的基因整合表达和Bacillus subtilis的拷贝数放大

Haoyu Guo^1,2,3,4, Rongzhen Tian^1,2,3,4, Yaokang Wu^1,2,3,4

¹Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China.

Synthetic and systems biotechnology

|May 6, 2024

概括

我们开发了BacAmp,Bacillus subtilis中稳定基因放大系统. 这种方法增强基因表达,改善生物生产和代谢工程应用.

科学领域:

合成生物学合成生物学
微生物工程微生物工程
遗传学是一个遗传学.

背景情况:

稳定,高水平的基因表达对于合成生物学中的微生物细胞工厂至关重要.
染色体基因放大对基于等离子体的系统具有优势,包括增强稳定性和降低代谢负担.
在Bacillus subtilis中扩大基因的现有方法在稳定性和效率上可能受到限制.

研究的目的:

开发一种用于稳定基因整合和Bacillus subtilis的副本数放大的新系统.
为控制基因组编辑和基因放大创建可逆基因开关.
为了证明该系统在增强基因表达和生物生产方面的有效性.

主要方法:

开发了使用可逆开关控制recA基因通过抑制剂和非天然氨基酸依赖表达的BacAmp系统.
激活了基因组编辑和基因放大开关,然后关闭它以稳定基因拷贝数.
利用绿色光蛋白 (GFP) 作为稳定基因放大标记.

主要成果:

通过基因放大,实现了基因表达的3倍增长,在110代后保持平均基因拷贝数为10个.
成功地应用了该系统来增强N-乙神经氨基酸 (NeuAc) 合成,将关键基因拷贝数增加到7.7并产生1.3倍的NeuAc标位.
证明了BacAmp系统对持续高水平基因表达的稳定性和有效性.

结论:

该BacAmp系统提供了一个强大的和稳定的方法,用于在Bacillus subtilis.
这项技术为合成生物学和代谢工程中的基因表达策略提供了一种新方法.
在微生物宿主中,BacAmp系统具有优化生物生产过程的巨大潜力.

关键词:

这种细菌是 Bacillus subtilis.同类的重组组合.稳定基因放大系统稳定基因放大系统收藏收藏收藏收藏收藏

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