Tetracycline-inducible expression systems with reduced basal activity in mammalian cells
Tetracycline-inducible expression systems with reduced basal activity in mammalian cells
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DOI：
10.1093/nar/27.2.708
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发表时间：
1999-01-15
影响因子：
14.9
通讯作者：
Hillen, W
中科院分区：
文献类型：
--
作者：
Forster, K;Helbl, V;Hillen, W
我们描述了四环素诱导的真核基因表达系统的修改，降低了HeLa细胞中的基础表达水平。它利用四环素诱导的反式激活因子和作用于同一启动子的四环素调节的阻遏因子融合。为了避免异源二聚化或竞争相同的DNA位点，每个DNA被提供不同的DNA识别和/或蛋白质二聚化特异性。我们在未诱导的状态下实现了主动沉默，导致添加四环素后基础转录水平降低了近6倍，基因表达激活了数百倍。
We describe a modification of the tetracycline-inducible eukaryotic gene expression system with decreased basal levels of expression in HeLa cells. It employs the tetracycline-inducible transactivator and a tetracycline regulated repressor fusion acting on the same promoter. To avoid heterodimerization or competition for the same DNA site, each was provided with different DNA recognition and/or protein dimerization specificities. We achieved active silencing in the uninduced state resulting in similar to 6-fold reduced levels of basal transcription and several hundred-fold activation of gene expression upon addition of tetracycline.