Mating-based Overexpression Library Screening in Yeast

This method can help answer key questions in the neurodegenerative disease field, such as what genetic factors and pathways regulate the toxicity of disease-associated proteins. The main advantage of this technique is that the highly efficient process of yeast mating is used to screen yeast models against a large collection of library genes. Though this method can provide insight into the toxicity of neurodegenerative disease proteins, it can also be applied to the study of other cellular processes in yeast, such as tolerance of other stress conditions.

Begin this procedure by aliquoting five microliters per well of plasmid DNA from an arrayed plasmid library to round bottom 96-well plates. Keep the plates at four degrees Celsius. Next, inoculate 150 milliliters of yeast peptone dextrose or YPD medium in a 500 milliliter flask with a colony of the haploid yeast strain W303 Alpha.