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

Ting Yuan, Yakun Guo, Junkai Dong, Tianyi Li, Tong Zhou, Kaiwen Sun, Mei Zhang, Qingyu Wu, Zhen Xie, Yizhi Cai, Limin Cao, Junbiao Dai*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

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.[Figure not available: see fulltext.]

Original languageEnglish
Pages (from-to)107-116
Number of pages10
JournalFrontiers of Chemical Science and Engineering
Volume11
Issue number1
Early online date22 Mar 2017
DOIs
Publication statusPublished - 22 Mar 2017

Keywords

  • metabolic engineering
  • promoter activity
  • synthetic biology
  • xylose utilization
  • yeast

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