A Layered Genetic Design Enables the Yeast Galactose Regulon to Respond to Cyanamide

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-...

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Bibliographic Details
Published in:ACS synthetic biology 2023-09, Vol.12 (9), p.2783-2788
Main Authors: Song, Jingya, Fan, Jian, Fan, Cong, He, Nike, Ye, Xixi, Cao, Mingfeng, Yuan, Jifeng
Format: Article
Language:English
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Summary: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.
ISSN:2161-5063
2161-5063
DOI:10.1021/acssynbio.3c00241