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)
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| 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. | ||||
