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Yeast. 2013 Sep;30(9):341-51. doi: 10.1002/yea.2967. Epub 2013 Jul 23.

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

Author information

1
Cancer Research UK London Research Institute, Clare Hall Laboratories, South Mimms, UK.

Abstract

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.

KEYWORDS:

auxin; budding yeast; degron; protein degradation; protein stability

PMID:
23836714
PMCID:
PMC4171812
DOI:
10.1002/yea.2967
[Indexed for MEDLINE]
Free PMC Article
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