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Gene Tagging Strategies To Assess Protein Expression, Localization, and Function in Drosophila
Analysis of gene function in complex organisms relies extensively on tools to detect the cellular and subcellular localization of gene products, especially proteins. Typically, immunostaining with antibodies provides these data. However, due to cost, time, and labor limitations, generating specific...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Genetics Society of America
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5629313/ https://www.ncbi.nlm.nih.gov/pubmed/28978772 http://dx.doi.org/10.1534/genetics.117.199968 |
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author | Kanca, Oguz Bellen, Hugo J. Schnorrer, Frank |
author_facet | Kanca, Oguz Bellen, Hugo J. Schnorrer, Frank |
author_sort | Kanca, Oguz |
collection | PubMed |
description | Analysis of gene function in complex organisms relies extensively on tools to detect the cellular and subcellular localization of gene products, especially proteins. Typically, immunostaining with antibodies provides these data. However, due to cost, time, and labor limitations, generating specific antibodies against all proteins of a complex organism is not feasible. Furthermore, antibodies do not enable live imaging studies of protein dynamics. Hence, tagging genes with standardized immunoepitopes or fluorescent tags that permit live imaging has become popular. Importantly, tagging genes present in large genomic clones or at their endogenous locus often reports proper expression, subcellular localization, and dynamics of the encoded protein. Moreover, these tagging approaches allow the generation of elegant protein removal strategies, standardization of visualization protocols, and permit protein interaction studies using mass spectrometry. Here, we summarize available genomic resources and techniques to tag genes and discuss relevant applications that are rarely, if at all, possible with antibodies. |
format | Online Article Text |
id | pubmed-5629313 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Genetics Society of America |
record_format | MEDLINE/PubMed |
spelling | pubmed-56293132017-10-18 Gene Tagging Strategies To Assess Protein Expression, Localization, and Function in Drosophila Kanca, Oguz Bellen, Hugo J. Schnorrer, Frank Genetics FlyBook Analysis of gene function in complex organisms relies extensively on tools to detect the cellular and subcellular localization of gene products, especially proteins. Typically, immunostaining with antibodies provides these data. However, due to cost, time, and labor limitations, generating specific antibodies against all proteins of a complex organism is not feasible. Furthermore, antibodies do not enable live imaging studies of protein dynamics. Hence, tagging genes with standardized immunoepitopes or fluorescent tags that permit live imaging has become popular. Importantly, tagging genes present in large genomic clones or at their endogenous locus often reports proper expression, subcellular localization, and dynamics of the encoded protein. Moreover, these tagging approaches allow the generation of elegant protein removal strategies, standardization of visualization protocols, and permit protein interaction studies using mass spectrometry. Here, we summarize available genomic resources and techniques to tag genes and discuss relevant applications that are rarely, if at all, possible with antibodies. Genetics Society of America 2017-10 2017-09-29 /pmc/articles/PMC5629313/ /pubmed/28978772 http://dx.doi.org/10.1534/genetics.117.199968 Text en Copyright © 2017 Kanca et al. Available freely online through the author-supported open access option. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | FlyBook Kanca, Oguz Bellen, Hugo J. Schnorrer, Frank Gene Tagging Strategies To Assess Protein Expression, Localization, and Function in Drosophila |
title | Gene Tagging Strategies To Assess Protein Expression, Localization, and Function in Drosophila |
title_full | Gene Tagging Strategies To Assess Protein Expression, Localization, and Function in Drosophila |
title_fullStr | Gene Tagging Strategies To Assess Protein Expression, Localization, and Function in Drosophila |
title_full_unstemmed | Gene Tagging Strategies To Assess Protein Expression, Localization, and Function in Drosophila |
title_short | Gene Tagging Strategies To Assess Protein Expression, Localization, and Function in Drosophila |
title_sort | gene tagging strategies to assess protein expression, localization, and function in drosophila |
topic | FlyBook |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5629313/ https://www.ncbi.nlm.nih.gov/pubmed/28978772 http://dx.doi.org/10.1534/genetics.117.199968 |
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