An expanded tool kit for the auxin‐inducible degron system in budding yeast |
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Authors: | Magdalena Morawska Helle D Ulrich |
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Affiliation: | 1. Cancer Research UK London Research Institute, Clare Hall Laboratories, , South Mimms, UK;2. Institute of Molecular Biology, , 55128 Mainz, Germany |
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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. © 2013 The Authors. Yeast published by John Wiley & Sons, Ltd. |
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Keywords: | degron auxin budding yeast protein degradation protein stability |
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