Construction, characterization and application of a genome-wide promoter library in Saccharomyces cerevisiae

Promoters are critical elements to control gene expression but could behave differently under various growth conditions. Here we report the construction of a genome-wide promoter library, in which each native promoter in Saccharomyces cerevisiae was cloned upstream of a yellow fluorescent protein (Y...

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Bibliographic Details
Published in:Frontiers of chemical science and engineering 2017-03, Vol.11 (1), p.107-116
Main Authors: Yuan, Ting, Guo, Yakun, Dong, Junkai, Li, Tianyi, Zhou, Tong, Sun, Kaiwen, Zhang, Mei, Wu, Qingyu, Xie, Zhen, Cai, Yizhi, Cao, Limin, Dai, Junbiao
Format: Article
Language:English
Subjects:
Bacteria
Baking yeast
Bins
Chemistry
Chemistry and Materials Science
Fermentation
Fluorescence
Gene expression
Genomes
Industrial Chemistry/Chemical Engineering
Libraries
Nanotechnology
Pools
Research Article
Xylose
Yeast
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Summary:Promoters are critical elements to control gene expression but could behave differently under various growth conditions. Here we report the construction of a genome-wide promoter library, in which each native promoter in Saccharomyces cerevisiae was cloned upstream of a yellow fluorescent protein (YFP) reporter gene. Nine libraries were arbitrarily defined and assembled in bacteria. The resulting pools of promoters could be prepared and transformed into a yeast strain either as centromeric plasmids or integrated into a genomic locus upon enzymatic treatment. Using fluorescence activated cell sorting, we classified the yeast strains based on YFP fluorescence intensity and arbitrarily divided the entire library into 12 bins, representing weak to strong promoters. Several strong promoters were identified from the most active bins and their activities were assayed under different growth conditions. Finally, these promoters were applied to drive the expression of genes in xylose utilization to improve fermentation efficiency. Together, this library could provide a quick solution to identify and utilize desired promoters under user-defined growth conditions.
ISSN:2095-0179
2095-0187
DOI:10.1007/s11705-017-1621-7