{"title":"Auxin-Inducible Degron System for Depletion of Proteins in Saccharomyces cerevisiae","authors":"Ameet Shetty, Natalia I. Reim, Fred Winston","doi":"10.1002/cpmb.104","DOIUrl":null,"url":null,"abstract":"<p>The auxin-inducible degron (AID) is a powerful tool that is used for depletion of proteins to study their function in vivo. This method can conditionally induce the degradation of any protein by the proteasome simply by the addition of the plant hormone auxin. This approach is particularly valuable to study the function of essential proteins. The protocols provided here describe the steps to construct the necessary strains and to optimize auxin-inducible depletion in <i>Saccharomyces cerevisiae</i>. © 2019 by John Wiley & Sons, Inc.</p><p><b>Basic Protocol 1</b>: Construction of TIR1-expressing strains by transformation</p><p><b>Basic Protocol 2</b>: Tagging a yeast protein of interest with an auxin-inducible degron</p><p><b>Support Protocol</b>: Construction of depletion strains by genetic crosses</p><p><b>Basic Protocol 3</b>: Optimization for depletion of the auxin-inducible-degron-tagged protein</p>","PeriodicalId":10734,"journal":{"name":"Current Protocols in Molecular Biology","volume":"128 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2019-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmb.104","citationCount":"10","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Molecular Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpmb.104","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 10
Abstract
The auxin-inducible degron (AID) is a powerful tool that is used for depletion of proteins to study their function in vivo. This method can conditionally induce the degradation of any protein by the proteasome simply by the addition of the plant hormone auxin. This approach is particularly valuable to study the function of essential proteins. The protocols provided here describe the steps to construct the necessary strains and to optimize auxin-inducible depletion in Saccharomyces cerevisiae. © 2019 by John Wiley & Sons, Inc.
Basic Protocol 1: Construction of TIR1-expressing strains by transformation
Basic Protocol 2: Tagging a yeast protein of interest with an auxin-inducible degron
Support Protocol: Construction of depletion strains by genetic crosses
Basic Protocol 3: Optimization for depletion of the auxin-inducible-degron-tagged protein
酿酒酵母蛋白质耗竭的生长素诱导脱菌系统
生长素诱导降解(AID)是研究蛋白质在体内功能的有力工具。该方法只需添加植物激素生长素，就能有条件地诱导蛋白酶体降解任何蛋白质。这种方法对于研究必需蛋白质的功能特别有价值。本文提供的方案描述了在酿酒酵母中构建必要菌株和优化生长素诱导耗尽的步骤。©2019 by John Wiley &基本方案1:通过转化构建表达tir1的菌株基本方案2:用生长素诱导的脱格标记感兴趣的酵母蛋白支持方案:通过遗传杂交构建耗尽菌株基本方案3:生长素诱导的脱格标记蛋白的耗尽优化
本文章由计算机程序翻译，如有差异，请以英文原文为准。
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