Single-chain Tet transregulators
Single-chain Tet transregulators
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DOI：
10.1093/nar/gkg421
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发表时间：
2003-06-15
影响因子：
14.9
通讯作者：
Hillen, W
中科院分区：
文献类型：
--
作者：
Krueger, C;Berens, C;Hillen, W
我们在此证明，Tet 阻遏蛋白 (TetR) 是一种二聚体变构调节蛋白，当通过 29 个氨基酸接头连接时，可以转化为全功能单体。基于 TetR 的反式调节子广泛用于调节真核生物中的基因表达。它们可以融合形成具有两个 TetR 部分和一个真核调节结构域的单链 (sc) Tet 反式调节子。反式激活子和反式沉默子的 Sc 变体与人类细胞系中各自的二聚体表现出相同的调节特性。特别是，rtTA 变体的反向“tet-on”表型也存在于 sc 变体中。逆反式激活子和 sc 反式沉默子的共表达导致背景表达减少，并在诱导后显示出完全激活。数据表明，sc Tet 反式调节子表现出各自二聚体的表型，并且在同一细胞中共表达时缺乏功能干扰。
We demonstrate here that the Tet repressor ( TetR), a dimeric allosterical regulatory protein, can be converted to a fully functional monomer when connected by a 29 amino acid linker. TetR-based transregulators are widely used to regulate gene expression in eukaryotes. They can be fused to form single-chain (sc) Tet transregulators with two TetR moieties and one eukaryotic regulatory domain. Sc variants of transactivator and transsilencer exhibit the same regulatory properties as their respective dimeric counterparts in human cell lines. In particular, the reverse 'tet-on' phenotype of rtTA variants is also present in the sc variants. Coexpression of a reverse transactivator and sc transsilencer leads to reduced background expression and shows full activation upon induction. The data demonstrate that sc Tet transregulators exhibit the phenotype of their respective dimers and lack functional interference when coexpressed in the same cell.