Isolation of a cDNA clone encoding a biologically active thyroid hormone receptor.
Isolation of a cDNA clone encoding a biologically active thyroid hormone receptor.
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分离编码具有生物活性的甲状腺激素受体的 cDNA 克隆。
DOI：
10.1073/pnas.85.14.5031
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发表时间：
1988
影响因子：
11.1
通讯作者：
Moore,DD
中科院分区：
文献类型：
--
作者：
Koenig,RJ;Warne,RL;Brent,GA;Harney,JW;Larsen,PR;Moore,DD
我们从GH3细胞文库中克隆了c-erba基因。该克隆命名为erb62，全长4.5kb，编码461个氨基酸的β型c-erba蛋白。这种c-erba蛋白与3，5，3‘-三碘甲状腺原氨酸(T3)及其类似物结合，亲和力类似于真正的T3受体。通过RNA凝胶印迹分析，erb62与在表达T3受体的器官中发现的6千碱基的RNA杂交，例如心脏、肾脏和大脑。COS-细胞瞬时共转染系统显示，erb62编码具有生物活性的T3受体。将对应于大鼠生长激素基因5‘侧翼区包含T3反应元件的一部分的寡核苷酸插入到氯霉素乙酰转移酶表达载体中单纯疱疹病毒胸苷激酶启动子的5’侧。该复合启动子引导的报告基因的表达只有在该质粒与表达erb62的质粒共转染时才能诱导表达。
We have isolated a c-erbA cDNA clone from a GH3 cell library. The clone, denoted erb62, is 4.5 kilobases long and encodes a 461-amino acid beta-type c-erbA protein. This c-erbA protein binds 3,5,3'-triiodothyronine (T3) and T3 analogs with affinities similar to those of the authentic T3 receptor. By RNA gel blot analysis, erb62 hybridizes to a 6-kilobase RNA found in organs that express T3 receptors--e.g., heart, kidney, and brain. A COS-cell transient cotransfection system was used to show that erb62 encodes a biologically active T3 receptor. An oligonucleotide, corresponding to a portion of the rat growth hormone gene 5'-flanking region that contains a T3 response element, was inserted on the 5' side of the herpes simplex virus thymidine kinase promoter in a chloramphenicol acetyltransferase-expressing plasmid. Reporter gene expression directed by this hybrid promoter was T3 inducible only if this plasmid was cotransfected with an erb62-expressing plasmid.