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An expanded tool kit for the auxin-inducible degron system in budding yeast
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 seri...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Blackwell Publishing Ltd
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4171812/ https://www.ncbi.nlm.nih.gov/pubmed/23836714 http://dx.doi.org/10.1002/yea.2967 |
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author | Morawska, Magdalena Ulrich, Helle D |
author_facet | Morawska, Magdalena Ulrich, Helle D |
author_sort | Morawska, Magdalena |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-4171812 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Blackwell Publishing Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-41718122014-10-08 An expanded tool kit for the auxin-inducible degron system in budding yeast Morawska, Magdalena Ulrich, Helle D Yeast Research Articles 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. Blackwell Publishing Ltd 2013-09 2013-07-23 /pmc/articles/PMC4171812/ /pubmed/23836714 http://dx.doi.org/10.1002/yea.2967 Text en Copyright © 2013 John Wiley & Sons, Ltd. http://creativecommons.org/licenses/by-nc/3.0/ This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. |
spellingShingle | Research Articles Morawska, Magdalena Ulrich, Helle D An expanded tool kit for the auxin-inducible degron system in budding yeast |
title | An expanded tool kit for the auxin-inducible degron system in budding yeast |
title_full | An expanded tool kit for the auxin-inducible degron system in budding yeast |
title_fullStr | An expanded tool kit for the auxin-inducible degron system in budding yeast |
title_full_unstemmed | An expanded tool kit for the auxin-inducible degron system in budding yeast |
title_short | An expanded tool kit for the auxin-inducible degron system in budding yeast |
title_sort | expanded tool kit for the auxin-inducible degron system in budding yeast |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4171812/ https://www.ncbi.nlm.nih.gov/pubmed/23836714 http://dx.doi.org/10.1002/yea.2967 |
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