An expanded tool kit for the auxin-inducible degron system in budding yeast

被引:282
作者
Morawska, Magdalena [1 ]
Ulrich, Helle D. [1 ,2 ]
机构
[1] Canc Res UK London Res Inst, Clare Hall Labs, S Mimms, Herts, England
[2] Inst Mol Biol, D-55128 Mainz, Germany
关键词
degron; auxin; budding yeast; protein degradation; protein stability; PROTEINS; GENE; REPLICATION; DEGRADATION; DEPLETION; DOMAIN;
D O I
10.1002/yea.2967
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Fusion of inducible degradation signals, so-called degrons, to cellular proteins is an elegant method of controlling protein levels in vivo. Recently, a degron system relying on the plant hormone auxin has been described for use in yeast and vertebrate cells. We now report the construction of a series of vectors that significantly enhance the versatility of this auxin-inducible degron (AID) system in Saccharomyces cerevisiae. We have minimized the size of the degron and appended a series of additional epitope tags, allowing detection by commercial antibodies or fluorescence microscopy. The vectors are compatible with PCR-based genomic tagging strategies, allow for C- or N-terminal fusion of the degron, and provide a range of selection markers. Application to a series of yeast proteins, including essential replication factors, provides evidence for a general usefulness of the system. (c) 2013 The Authors. Yeast published by John Wiley & Sons, Ltd.
引用
收藏
页码:341 / 351
页数:11
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