The creation of a genome-wide yeast deletion collection revealed that there are over 1,000 protein-coding genes that are essential for yeast viability. The function of many of these essential genes has yet to be defined. In part, this is due to the difficulty of utilizing essential genes in functional assays. The Tet-Promoter Hughes Collection (yTHC) from the Hughes Laboratory, University of Toronto, helps to overcome this barrier.
The yTHC collection contains 800 essential yeast genes for which expression is regulated by doxycycline. The endogenous promoter has been replaced with a Tet-titratable promoter in the genome. This allows the expression of the gene to be switched off by the addition of doxycycline to the yeast growth medium.
Yeast Tet-Promoters Hughes Collection (yTHC) mutant strains are provided in the haploid MATa strain R1158
Each strain was qualified by its growth phenotype on YPD with 10 µg/mL doxycycline
In the absence of doxycycline, the Tet-promoter is fully activated
Addition of doxycycline in a titratable manner allows for down-regulation of the promoter until the gene of interest is no longer expressed at detectable levels
Background Strain = R1158 (derived from By4741)
Genotype of wildtype R1158 - URA3::CMV-tTA MATa his3-1 leu2-0 met15-0
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Wildtype R1158 is provided in YPD with addition of 15% glycerol following incubation.