Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cells
Summary: 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 overexpress...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722006657 |
| _version_ | 1811292003017687040 |
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| author | Ana Luiza Drumond-Bock Magdalena Cybula Luyao Wang Lin Wang Magdalena Bieniasz |
| author_facet | Ana Luiza Drumond-Bock Magdalena Cybula Luyao Wang Lin Wang Magdalena Bieniasz |
| author_sort | Ana Luiza Drumond-Bock |
| collection | DOAJ |
| description | Summary: 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. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-13T04:38:55Z |
| format | Article |
| id | doaj.art-8599d499c2dd4291ad6fee99695de249 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-13T04:38:55Z |
| publishDate | 2022-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-8599d499c2dd4291ad6fee99695de2492022-12-22T03:02:05ZengElsevierSTAR Protocols2666-16672022-12-0134101785Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cellsAna Luiza Drumond-Bock0Magdalena Cybula1Luyao Wang2Lin Wang3Magdalena Bieniasz4Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA; Corresponding authorCytovance® Biologics, Oklahoma City, OK 73104, USAOklahoma Medical Research Foundation, Oklahoma City, OK 73104, USAOklahoma Medical Research Foundation, Oklahoma City, OK 73104, USAOklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA; Corresponding authorSummary: 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. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166722006657Cell biologyMolecular biologyProtein biochemistryProtein expression and purification |
| spellingShingle | Ana Luiza Drumond-Bock Magdalena Cybula Luyao Wang Lin Wang Magdalena Bieniasz Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cells STAR Protocols Cell biology Molecular biology Protein biochemistry Protein expression and purification |
| 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 | Cell biology Molecular biology Protein biochemistry Protein expression and purification |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722006657 |
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