在高效率和精度下构建和多样化天然产品生物合成基因集群
Chaoxian Bai1, Lina M Bayona1, Gilles P van Wezel1
1Institute of Biology, Leiden University, Sylviusweg 72, 2333 BE Leiden, Netherlands.
ACS synthetic biology
|October 10, 2025
概括
我们开发了一种金门组装 (GGA) 方法,用于准确地克隆和工程生物合成基因集群 (BGCs). 这个平台加速了用于药物开发的新型天然产品的发现.
科学领域:
- 合成生物学 合成生物学
- 基因组学就是基因组学.
- 自然产品的生物合成.
背景情况:
- 生物合成基因集群 (BGCs) 对于生产用于治疗的天然产品至关重要.
- 由于表达挑战,许多BGC没有特征,阻碍了药物发现.
- 现有的克隆方法,如TAR和吉布森组装,容易出现错误.
研究的目的:
- 提出一种新的,准确的,有效的策略,用于BGC克隆和重构使用等级的金门组合 (GGA).
- 为了实现系统的途径工程和基因组基因组药物发现的突变发生.
主要方法:
- 开发了一个基于Golden Gate Assembly (GGA) 的平台,用于BGC的构建和重构.
- 设计了23kb的actinorhodin (ACT) BGC和23种突变衍生物,具有高效率.
- 在*Streptomyces coelicolor* M1152中表达的重构BGC和使用GNPS分子网络分析的产品.
主要成果:
- 在构建ACT BGC及其突变物时实现了100%的效率.
- 确定了九个ACT产生必需的基因,并观察了基因失活时的途径重新连接.
- 发现了许多未知的分子,扩大了已知的ACT生物合成的化学空间.
结论:
- 建立了一个强大的基于GGA的平台,用于BGC的构建,重构和功能剖析.
- 该平台加速合成生物学驱动的天然产品发现.
- 证明了GGA在精确基因操纵和探索新型化学实体方面的实用性.
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