Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cells.
Cloning BRD4 long isoform into overexpression vectors for stable overexpression of BRD4-L in mammalian cells.
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DOI：
10.1016/j.xpro.2022.101785
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发表时间：
2022-12-16
期刊：
影响因子：
--
通讯作者：
Bieniasz M
中科院分区：
文献类型：
--
作者：
Drumond-Bock AL;Cybula M;Wang L;Wang L;Bieniasz M
BRD 4-L的分子克隆是一项具有挑战性的技术，因为DNA插入物是由一个长的富含GC的序列形成的，该序列折叠成二级结构。本方案定义了扩增BRD 4-L的特定策略，随后将基因成功克隆到过表达载体中。由于没有现有的方案，也没有商业上可用的质粒，这项工作提供了一个有用的工具，涉及BRD 4-L的分子克隆的研究，并可能被应用到其他具有挑战性的基因。PCR扩增富含GC的长序列BRD 4-L与过表达载体LentiV_Blast的连接质粒LentiV_Blast-BRD 4-L的构建和测序出版商说明：进行任何实验方案都需要遵守当地机构的实验室安全和伦理指南。BRD 4-L的分子克隆是一项具有挑战性的技术，因为DNA插入物是由一个长的富含GC的序列形成的，该序列折叠成二级结构。本方案定义了扩增BRD 4-L的特定策略，随后将基因成功克隆到过表达载体中。由于没有现有的方案，也没有商业上可用的质粒，这项工作提供了一个有用的工具，涉及BRD 4-L的分子克隆的研究，并可能被应用到其他具有挑战性的基因。
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. PCR amplification of a long, GC-rich sequence Ligation of BRD4-L with the overexpression vector LentiV_Blast Construction and sequencing of the plasmid LentiV_Blast-BRD4-L Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.
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影响因子：
14.9
作者：
Kibbe WA
通讯作者：
Kibbe WA
影响因子：
64.8
作者：
Shu S;Lin CY;He HH;Witwicki RM;Tabassum DP;Roberts JM;Janiszewska M;Huh SJ;Liang Y;Ryan J;Doherty E;Mohammed H;Guo H;Stover DG;Ekram MB;Brown J;D'Santos C;Krop IE;Dillon D;McKeown M;Ott C;Qi J;Ni M;Rao PK;Duarte M;Wu SY;Chiang CM;Anders L;Young RA;Winer E;Letai A;Barry WT;Carroll JS;Long H;Brown M;Liu XS;Meyer CA;Bradner JE;Polyak K
通讯作者：
Polyak K
DOI：
10.1158/1078-0432.ccr-14-3283
发表时间：
2015-12-15
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Bieniasz M;Radhakrishnan P;Faham N;De La O JP;Welm AL
通讯作者：
Welm AL
影响因子：
3.1
作者：
Mamedov, T. G.;Pienaar, E.;Whitney, S. E.;TerMaat, J. R.;Carvill, G.;Goliath, R.;Subramanian, A.;Viljoen, H. J.
通讯作者：
Viljoen, H. J.
影响因子：
--
作者：
Green, Michael R;Sambrook, Joseph
通讯作者：
Sambrook, Joseph