Experiment Overview

Repository ID: FR-FCM-ZY8Y Experiment name: Activity of 48 TDH3 promoter variants in YPD MIFlowCyt score: 49.25%
Primary researcher: Fabien Duveau PI/manager: Patricia Wittkopp Uploaded by: Fabien Duveau
Experiment dates: 2016-04-02 - 2016-04-02 Dataset uploaded: Jul 2017 Last updated: Aug 2017
Keywords: [cis-regulatory mutations] [expression noise] [promoter activity] [fitness] Manuscripts:
Organizations: University of Michigan, Ann Arbor, MI (USA)
Purpose: Quantifying the effects of 48 alleles of the TDH3 promoter on the mean activity and the variability of activity (noise) of the promoter. These promoter variants were cloned upstream of a YFP reporter gene and inserted at the HO locus in the yeast genome. Most alleles carry mutations located in previously characterized transcription factor binding sites or/and in the TATA box that were selected to cover a broad range of mean and noise of activity. Some alleles contain two copies of the TDH3 promoter, used to obtain achieve overexpression of the reporter gene. The same alleles of the TDH3 promoter were also cloned upstream of the native TDH3 coding sequence to quantify their effect on fitness in another experiment.
Conclusion: The mean activity varied from 0.1% to 207% of the wild type level among the 48 alleles of the TDH3 promoter. Standard deviation of activity varied from 1.6% to 448% of wild type standard deviation among alleles.
Comments: None
Funding: Not disclosed
Quality control: Samples were grown and scored in four replicate 96 well plates, for a total of 384 samples. Each plate contained 24 replicates of a control strain containing a wild type allele of the TDH3 promoter driving expression of YFP. These controls can be used to correct for technical variation between plates or flow cytrometry runs.
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