Experiment Overview

Repository ID: FR-FCM-Z3J3 Experiment name: Expression of pTDH3-YFP in GCR1 mutant strains of yeast MIFlowCyt score: 24.75%
Primary researcher: Fabien Duveau PI/manager: Patricia Wittkopp Uploaded by: Fabien Duveau
Experiment dates: 2017-01-24 - 2017-01-25 Dataset uploaded: Mar 2021 Last updated: Mar 2021
Keywords: None Manuscripts:
Organizations: University of Michigan, Ann Arbor, MI (USA)
Purpose: The goal of this experiment is to quantify the effects of random mutations in the coding sequence of the GCR1 gene on expression of the reporter gene pTDH3-YFP (YFP expressed under control of the TDH3 promoter in yeast). GCR1 encodes a transcription factor known to directly bind the TDH3 promoter. The experiment includes 4 replicate populations of 683 strains with random mutations in GCR1 sequence. For each sample, the fluorescence of at least 1000 cells was measured.
Conclusion: Random mutations in GCR1 are likely to alter the activity of the TDH3 promoter (38% of mutants showed a decreased pTDH3-YFP expression by more than 3%). Null alleles of GCR1 decrease expression of pTDH3-YFP by 80% relative to a strain with wild type GCR1 sequence.
Comments: None
Funding: Not disclosed
Quality control: The experiment includes 96 replicate populations of a strain that does not contain the pTDH3-YFP reporter gene and that is used to estimate the level of autofluorescence. 204 samples correspond to replicate populations of the wild-type progenitor strain used as a reference to quantify expression changes in each mutant. An additional 24 wild-type strains expressing pTDH3-YFP were arrayed at predetermined positions in each 96-well plates to correct for technical variation between plates or flow cytometry runs.
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