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

ACS Synth Biol

State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University, Fujian 361102, China.

Published: September 2023

The commonly used expression systems in typically rely on either constitutive or galactose-regulated promoters. The lack of inducible systems in 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.

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Source
http://dx.doi.org/10.1021/acssynbio.3c00241DOI Listing

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