Expression of pTDH3-YFP in trans-regulatory mutant strains of yeast

The goal of this experiment is to compare the expression level of the pTDH3-YFP reporter gene (YFP expressed under control of the TDH3 yeast promoter) between EMS mutants and single-site mutants that only contain a single mutation expected to alter fluorescence from genetic mapping experiments. For each genotype, fluorescence was quantified from 4 replicate populations cultivated in separate wells of 96-well plates at two different dates (8 replicate populations in total). For each replicate population, at least 1000 cells were analyzed by flow cytometry.
Conclusion:
A strong correlation of pTDH3-YFP expression level is observed between EMS mutants and single-site mutants sharing the same mutation. The experiment includes several replicate populations of a strain that does not contain the pTDH3-YFP reporter gene and that is used to estimate the level of autofluorescence. 56 samples correspond to replicate populations of the wild-type progenitor strain used as a reference to quantify the changes of expression 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.