MICR 302 Lecture Notes - Lecture 4: Phenotypic Screening, Galactose, His3
7 Apr 2018
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Example: insert 21 genes into a yeast strain, knockout 3 = generate strictosidine in yeast (normally derived from plants) that is normally expensive. We know the use of forward selection marker, where yeast growth requires expression of the gene (ura3, his3, leu2, g418r) Counter selectable markers work in the opposite way and can be used to select against and kill the cell that expresses the counter selectable marker. Studying mutants of essential genes: need to conditionally perturb the function of essential genes since knocking them out will kill the cell. In a cell that is yfg1:nat: insert a wt yfg1 on a plasmid with ura3. Now you have a cell with both the fully functional plasmid yfg1 on. Ura3 and a mutated allele of yfg1 on a plasmid with his3 genes. Exposing the cells of 5-foa: you would expect growth of cells with the mutated yfg1 on his3.
