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Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cells
Molecular cloning of BRD4-L is a challenging technique, because the DNA insert is formed by a long, GC-rich sequence, which folds into secondary structures. The present protocol defines a specific strategy to amplify BRD4-L, followed by the successful cloning of the gene into an overexpression vecto...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9617197/ https://www.ncbi.nlm.nih.gov/pubmed/36317179 http://dx.doi.org/10.1016/j.xpro.2022.101785 |
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author | Drumond-Bock, Ana Luiza Cybula, Magdalena Wang, Luyao Wang, Lin Bieniasz, Magdalena |
author_facet | Drumond-Bock, Ana Luiza Cybula, Magdalena Wang, Luyao Wang, Lin Bieniasz, Magdalena |
author_sort | Drumond-Bock, Ana Luiza |
collection | PubMed |
description | Molecular cloning of BRD4-L is a challenging technique, because the DNA insert is formed by a long, GC-rich sequence, which folds into secondary structures. The present protocol defines a specific strategy to amplify BRD4-L, followed by the successful cloning of the gene into an overexpression vector. Since there are no existing protocols nor commercially available plasmids, this work provides a useful tool for studies involving molecular cloning of BRD4-L and could potentially be applied to other challenging genes. |
format | Online Article Text |
id | pubmed-9617197 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-96171972022-10-30 Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cells Drumond-Bock, Ana Luiza Cybula, Magdalena Wang, Luyao Wang, Lin Bieniasz, Magdalena STAR Protoc Protocol Molecular cloning of BRD4-L is a challenging technique, because the DNA insert is formed by a long, GC-rich sequence, which folds into secondary structures. The present protocol defines a specific strategy to amplify BRD4-L, followed by the successful cloning of the gene into an overexpression vector. Since there are no existing protocols nor commercially available plasmids, this work provides a useful tool for studies involving molecular cloning of BRD4-L and could potentially be applied to other challenging genes. Elsevier 2022-10-26 /pmc/articles/PMC9617197/ /pubmed/36317179 http://dx.doi.org/10.1016/j.xpro.2022.101785 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Drumond-Bock, Ana Luiza Cybula, Magdalena Wang, Luyao Wang, Lin Bieniasz, Magdalena Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cells |
title | Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cells |
title_full | Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cells |
title_fullStr | Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cells |
title_full_unstemmed | Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cells |
title_short | Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cells |
title_sort | cloning brd4 long isoform into overexpression vectors for stable overexpression of brd4-l in mammalian cells |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9617197/ https://www.ncbi.nlm.nih.gov/pubmed/36317179 http://dx.doi.org/10.1016/j.xpro.2022.101785 |
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