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一个严格可诱导和正交的Dre-rox系统,用于精确和无标记的基因组编辑Bacillus subtilis

Jianan Lv^1,2,3, Gang Fu^2,3, Qiyao Zhu^2,3

¹School of Biological Engineering, Dalian Polytechnic University, Dalian 116034, P.R. China.

Journal of microbiology and biotechnology

|July 29, 2025

概括

我们优化了Dre-rox系统,用于精确的基因组工程 Bacillus subtilis. 这种直角位点特定的重组系统使得 prokaryotes 中的模块化工作流和多位点工程成为可能.

科学领域:

合成生物学合成生物学
微生物基因组学微生物基因组学
分子生物学分子生物学

背景情况:

特定位置重组 (SSR) 对于微生物精确的基因组工程至关重要.
克雷复合酶在Bacillus subtilis中被广泛使用,但需要正交的系统.
开发新的SSR扩大了基因组工程工具包.

研究的目的:

建立和优化Dre-rox系统用于Bacillus subtilis的基因组工程.
为了评估Dre-rox与Cre-rox相比的正交性和效率.
为了证明Dre-rox对模块化和多位置基因组修改的实用性.

主要方法:

在B. subtilis.中利用了基于抗性基因的记者系统.
采用一种可诱导神氨酸的核糖开关来控制Dre表达并最大限度地减少泄漏的重组.
经过测试的CRE介导整合,其次是Dre介导无标记删除.
验证了使用双部位和三部位模型的同步多部位切除.

主要成果:

德雷-罗克斯系统展示了高可靠性,直角重组,没有与Cre.
优化的Dre-rox架构确保了可靠的基因组工程,具有低毒性和高宿主适应性.
该系统实现了一个模块化工作流程,用于Cre介导的集成和Dre介导的无标记删除.
通过使用Dre-rox.的单次诱导实现了同步的多部位切除.

结论:

德雷-罗克斯系统是一个可靠和高效的工具,用于 prokaryotic 基因组工程.
这种直角的SSR平台促进了分层路径工程和模块化底盘开发.
该研究将Dre-rox的实用性扩展到微生物系统,增强合成生物学应用.

关键词:

这种细菌是 Bacillus subtilis.这是Dre-rox系统.调节RNA开关的调节没有标记的编辑.正角复合酶系统的正角复合酶系统.特定地点的重组组合.

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