论文部分内容阅读
利用体外翻译系统，翻译了能与雌激素效应元件（ＥＲＥ）结合的全长的人雌激素受体（ｈＥＲ）。制备了切除卵巢的雌性大鼠子宫核抽提物，在雌激素存在下，此核抽提物能增强ｈＥＲ与ＥＲＥ的结合。此核抽提物在５０℃保温１５ｍｉｎ后，明显减弱了增强ｈＥＲ－ＥＲＥ结合的作用。提示了核抽提物中存在着能增强ｈＥＲ－ＥＲＥ结合的雌激素依赖的热敏感性的辅助因子。在大肠杆菌中表达了谷胱甘肽转硫酶（ＧＳＴ）融合的雌激素受体的ＤＮＡ结合区（ＥＲＤＢＤ），表达产物（ＧＳＴ－ＥＲＤＢＤ）也能与ＥＲＥ结合，但结合不受核抽提物影响。提示了核因子增强ｈＥＲ－ＥＲＥ结合的作用很可能不是通过ＤＢＤ的区域起作用的。
Using an in vitro translation system, the full-length human estrogen receptor (hER) that binds to the estrogen responsive element (ERE) is translated. An ovariectomized female rat uterine nuclear extract was prepared which, in the presence of estrogen, enhanced the binding of hER to ERE. After this nuclear extract was incubated at 50 ° C for 15 min, the effect of enhancing the binding of hER-ERE was significantly weakened. Suggesting that there is an estrogen-dependent thermostable cofactor in nuclear extracts that enhances hER-ERE binding. The DNA binding domain (ERDBD) of the estrogen receptor fused with glutathione S-transferase (GST) was expressed in E. coli and the expression product (GST-ERDBD) was also able to bind to ERE, but the binding was not inhibited by nuclear extraction Material effects. Suggesting that the effect of nuclear factor-enhanced hER-ERE binding probably does not work through the area of DBD.