一个用于真菌多基因通路的多重和预校准表达的多基因系统
1Biotechnology Research Institute, The Chinese Academy of Agricultural Sciences, Beijing, China.
Nature communications
|July 17, 2023
概括
研究人员开发了HACKing,这是一个新的系统,用于在真菌中有效地共同表达多个基因. 这种合成生物学工具能够精确控制代谢工程和代谢物生产的基因表达.
科学领域:
- 合成生物学 合成生物学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 多个基因的共同表达对于合成生物学应用至关重要.
- 现有的多基因路径组装方法可能是低效和复杂的.
- 需要对基因表达水平进行可预测的控制,以优化代谢途径.
研究的目的:
- 发现一种新的核酸序列,用于在酵母和丝状真菌中高效的多基因基因表达.
- 使用CRISPR/Cas9基因组编辑组装多基因通路的简化策略.
- 为了实现酶表达水平的预校准,以优化代谢物生产.
主要方法:
- 识别了一种9-bp序列,使其能够实现多基斯特龙表达.
- 开发HACKing (高效和可访问的系统通过CracKing基因进入基因组) 战略.
- 整合多基斯特隆表达与多重化,无标记CRISPR/Cas9基因组编辑.
- 将基因翻译链接到具有已知丰度的宿主蛋白质,用于表达校准.
主要成果:
- 在酵母和丝状真菌中证明了高效的多基因基因表达.
- 通过HACKing策略成功组装了多基因通路.
- 构建了Saccharomyces cerevisiae细胞工厂,表达了13个生物合成基因.
- 取得了大量的内源性烯 (1,090.41毫克L-1) 和异质性摩格罗尔 (1.04毫克L-1) 的生产.
结论:
- HACKing提供了一种可预测,简单,可扩展和快速的真菌通路工程方法.
- 该系统通过精确控制基因表达来优化有价值的代谢物生产.
- 这种方法提升了合成生物学在工业应用中的能力.
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