Rapid Depletion of Budding Yeast Proteins via the Fusion of an Auxin‐Inducible Degron (AID)

作者：Kohei Nishimura, Masato T. Kanemaki · 发表于：Current Protocols in Cell Biology · 年份：2014 · DOI：10.1002/0471143030.cb2009s64 · 被引用次数：88 · 研究领域：Plant Reproductive Biology、Transgenic Plants and Applications、Plant Molecular Biology Research

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.