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- W3083349616 abstract "Conditional mutants, such as temperature-sensitive (ts) mutants, are effective tools for the analysis of essential genes. However, such mutants are frequently leaky. To overcome this problem, it is helpful to isolate a tight conditional mutant of a gene of interest, e.g., by using ubiquitin-mediated protein degradation to eliminate the gene product. One such strategy is the auxin-inducible degron (AID) system, which is easy to use because the simple addition of auxin can induce the degradation of a target protein. Sometimes, however, elimination of the target protein is not sufficient, and an AID mutant exhibits a leaky phenotype. To address this issue, the improved AID (iAID) system was developed. In this approach, transcriptional repression by the promoter is combined with proteolytic elimination of the target protein by the AID system, yielding a much tighter mutant. Because simple addition of tetracycline is sufficient to repress the Tet-OFF promoter, the combination of Tet-OFF and AID maintains the ease of use of the original AID system. In this manuscript, we describe how to construct and use iAID mutants in the budding yeast Saccharomyces cerevisiae." @default.
- W3083349616 created "2020-09-11" @default.
- W3083349616 creator A5081256427 @default.
- W3083349616 date "2020-09-06" @default.
- W3083349616 modified "2023-09-26" @default.
- W3083349616 title "Construction of Tight Conditional Mutants Using the Improved Auxin-Inducible Degron (iAID) Method in the Budding Yeast Saccharomyces cerevisiae" @default.
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- W3083349616 doi "https://doi.org/10.1007/978-1-0716-0868-5_2" @default.
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