在哺乳动物细胞中具有降低基础活性的四环素诱导表达系统。
Tetracycline-inducible expression systems with reduced basal activity in mammalian cells.
作者信息
Forster K, Helbl V, Lederer T, Urlinger S, Wittenburg N, Hillen W
机构信息
Lehrstuhl für Mikrobiologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Staudtstrasse 5, D-91058 Erlangen, Germany.
出版信息
Nucleic Acids Res. 1999 Jan 15;27(2):708-10. doi: 10.1093/nar/27.2.708.
Abstract
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 approximately 6-fold reduced levels of basal transcription and several hundred-fold activation of gene expression upon addition of tetracycline.
摘要
我们描述了一种四环素诱导型真核基因表达系统的改进方法，该方法可降低HeLa细胞中的基础表达水平。它采用四环素诱导型反式激活因子和作用于同一启动子的四环素调节阻遏物融合体。为避免异源二聚化或对同一DNA位点的竞争，每个都具有不同的DNA识别和/或蛋白质二聚化特异性。我们在未诱导状态下实现了活性沉默，导致基础转录水平降低约6倍，添加四环素后基因表达激活数百倍。
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