title: Rapid Depletion of Budding Yeast Proteins via the Fusion of an Auxin-Inducible Degron (AID).
abstract: The auxin-inducible degron (AID) system allows the rapid and reversible proteolysis of proteins of interest, and enables the generation of conditional mutants of budding yeast. The construction of budding yeast AID mutants is simple, and the effect of depletion of essential proteins on proliferation can be confirmed by analyzing their phenotype. In this protocol, we describe a procedure to generate AID mutants of budding yeast via a simple transformation using PCR-amplified DNA. We also describe methods to confirm the depletion of proteins of interest that are required for proliferation by serial-dilution and liquid-culture assays.
authors.affiliation: Center of Frontier Research, National Institute of Genetics, Research Organization of Information and Systems, Shizuoka, Japan.
authors.forename: Kohei
authors.initials: K
authors.lastname: Nishimura
authors.forename: Masato T
authors.initials: MT
authors.lastname: Kanemaki
mesh_terms: D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D007210:Indoleacetic Acids / Q000494:pharmacology*; D009154:Mutation; D059748:Proteolysis; D004718:Saccharomycetales / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D005656:Fungal Proteins; D007210:Indoleacetic Acids
keywords: auxin; conditional mutant; degron; protein degradation; ubiquitin-proteasome pathway