Centre for Synthetic Biology and Innovation, Imperial College London, London, United Kingdom.
Department of Bioengineering, Imperial College London, London, United Kingdom.
PLoS One. 2018 Mar 22;13(3):e0194588. doi: 10.1371/journal.pone.0194588. eCollection 2018.
Engineered promoters with predefined regulation are a key tool for synthetic biology that enable expression on demand and provide the logic for genetic circuits. To expand the availability of synthetic biology tools for S. cerevisiae yeast, we here used hybrid promoter engineering to construct tightly-controlled, externally-inducible promoters that only express in haploid mother cells that have contributed a daughter cell to the population. This is achieved by combining elements from the native HO promoter and from a TetR-repressible synthetic promoter, with the performance of these promoters characterized by both flow cytometry and microfluidics-based fluorescence microscopy. These new engineered promoters are provided as an enabling tool for future synthetic biology applications that seek to exploit differentiation within a yeast population.
工程化的具有预设调控的启动子是合成生物学的关键工具，它可以实现按需表达，并为遗传回路提供逻辑。为了扩大合成生物学工具在酿酒酵母中的可用性，我们使用杂交启动子工程来构建严格控制的、可外部诱导的启动子，这些启动子只在已经向群体中贡献了一个子细胞的单倍体母细胞中表达。这是通过将天然 HO 启动子和 TetR 可抑制的合成启动子的元件结合来实现的，这些启动子的性能通过流式细胞术和基于微流控的荧光显微镜进行了表征。这些新的工程化启动子为未来寻求利用酵母群体内分化的合成生物学应用提供了一个使能工具。
