A reliable way of obtaining stable inducible clones
A reliable way of obtaining stable inducible clones
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DOI：
10.1093/nar/26.14.3443
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发表时间：
1998-07-15
影响因子：
14.9
通讯作者：
Pognonec, P
中科院分区：
文献类型：
--
作者：
Chambard, JC;Pognonec, P
诱导型基因表达系统为基因产物功能的分析提供了强有力的工具。四环素（Tc）表达系统已广泛且成功地应用于许多情况。然而，该系统的使用仍然有些繁琐，这是由于：（i）建立组成型和稳定表达Tc调节的反式激活因子的原代细胞系和（ii）获得以Tc依赖性方式表达目的基因的次级细胞系。为了促进这两个关键步骤，我们设计了一个有效的和分子生物学的自由的策略，允许成功选择克隆表达任何cDNA下严格的监管。
Inducible gene expression systems provide a powerful tool for the analysis of gene product functions. The 'Tetracycline (Tc) expression system' has been widely and successfully used in many instances. However, this system remains somewhat tedious to use due to: (i) the establishment of a primary cell line constitutively and stably expressing the Tc-regulated transactivator and (ii) the obtention of a secondary line expressing the gene of interest in a Tc-dependent manner. In order to facilitate these two critical steps, we devised an efficient and molecular biology-free strategy allowing the successful selection of clones expressing any cDNA under tight regulation.