Tetracycline-controlled transcription in eukaryotes: Novel transactivators with graded transactivation potential
Tetracycline-controlled transcription in eukaryotes: Novel transactivators with graded transactivation potential
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DOI：
10.1093/nar/25.14.2723
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发表时间：
1997-07-15
影响因子：
14.9
通讯作者：
Bujard, H
中科院分区：
文献类型：
--
作者：
Baron, U;Gossen, M;Bujard, H
产生了几种四环素控制的反式激活因子（tTA），它们的激活潜力相差>3个数量级。反式激活因子是泰特阻遏物和来源于单纯疱疹病毒蛋白16（VP16）的最小转录激活结构域之间的融合物。通过将先前描述的tTA的VP16部分减少至12个氨基酸，消除了与各种细胞转录因子相互作用的潜在靶标，以及可能引发细胞免疫应答的潜在表位。当与最初描述的tTA相比时，这些新的反式激活因子在更高的细胞内浓度下耐受。这将有助于在各种条件下建立泰特调控系统，但特别是当细胞类型限制的四环素控制的基因表达将通过同源重组在转基因生物中实现时。
Several tetracycline-controlled transactivators (tTA) were generated which differ in their activation potential by >3 orders of magnitude. The transactivators are fusions between the Tet repressor and minimal transcriptional activation domains derived from Herpes simplex virus protein 16 (VP16). By reducing the VP16 moiety of the previously described tTA to 12 amino acids, potential targets for interactions with various cellular transcription factors were eliminated, as were potential epitopes which may elicit a cellular immune response. When compared with the originally described tTA, these new transactivators are tolerated at higher intracellular concentrations. This will facilitate establishment of tet regulatory systems under a variety of conditions, but particularly when cell type-restricted tetracycline-controlled gene expression is to be achieved in transgenic organisms via homologous recombination.