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A Layered Genetic Design Enables the Yeast Galactose Regulon to Respond to Cyanamide
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2023-08-21 , DOI: 10.1021/acssynbio.3c00241
Jingya Song 1 , Jian Fan 1 , Cong Fan 1 , Nike He 1 , Xixi Ye 1 , Mingfeng Cao 2 , Jifeng Yuan 1
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2023-08-21 , DOI: 10.1021/acssynbio.3c00241
Jingya Song 1 , Jian Fan 1 , Cong Fan 1 , Nike He 1 , Xixi Ye 1 , Mingfeng Cao 2 , Jifeng Yuan 1
Affiliation
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The commonly used expression systems in Saccharomyces cerevisiae typically rely on either constitutive or galactose-regulated promoters. The lack of inducible systems in S. cerevisiae limits the precise temporal regulation of protein function and yeast metabolism. We herein repurposed the galactose-regulated system to make it respond to cyanamide. By using a cyanamide-inducible DDI2 promoter to control Gal4 expression in CEN.PK2–1C with Δgal80, a tight and graded cyanamide-inducible GAL system with an enhanced signal output was constructed. Subsequently, we demonstrated that the cyanamide-inducible GAL system was capable of tightly regulating the pentafunctional Aro1 protein to achieve conditional shikimate pathway activity. Taken together, the cyanamide-inducible GAL system could be implemented for both fundamental research and applied biotechnology.
中文翻译:
分层遗传设计使酵母半乳糖调节子能够响应氰胺
酿酒酵母中常用的表达系统通常依赖于组成型启动子或半乳糖调节启动子。酿酒酵母缺乏诱导系统限制了蛋白质功能和酵母代谢的精确时间调节。我们在此重新调整半乳糖调节系统的用途,使其对氰胺做出反应。通过使用氰胺诱导型 DDI2 启动子通过 Δgal80 控制 CEN.PK2–1C 中 Gal4 的表达,构建了具有增强信号输出的紧密且分级的氰胺诱导型 GAL 系统。随后,我们证明氰胺诱导的 GAL 系统能够严格调节五功能 Aro1 蛋白以实现条件莽草酸途径活性。总而言之,氰胺诱导的 GAL 系统可用于基础研究和应用生物技术。
更新日期:2023-08-21
中文翻译:
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分层遗传设计使酵母半乳糖调节子能够响应氰胺
酿酒酵母中常用的表达系统通常依赖于组成型启动子或半乳糖调节启动子。酿酒酵母缺乏诱导系统限制了蛋白质功能和酵母代谢的精确时间调节。我们在此重新调整半乳糖调节系统的用途,使其对氰胺做出反应。通过使用氰胺诱导型 DDI2 启动子通过 Δgal80 控制 CEN.PK2–1C 中 Gal4 的表达,构建了具有增强信号输出的紧密且分级的氰胺诱导型 GAL 系统。随后,我们证明氰胺诱导的 GAL 系统能够严格调节五功能 Aro1 蛋白以实现条件莽草酸途径活性。总而言之,氰胺诱导的 GAL 系统可用于基础研究和应用生物技术。