论文部分内容阅读
将雄激素受体(AR)的cDNA1119bp片段(1105~2224)(其中包含其DNA结合结构域到部分激素结合结构域)克隆于表达质粒pGEX中,通过IPTG诱导,在E.coli中表达GST-AR融合蛋白,经谷胱甘肽-Sepharose-4B亲和层析得以部分纯化。利用一个有雄激素应答元件(ARE)活性的C3(1)DNA片段为阳性探针,通过凝胶阻滞分析和DNaseI足迹法证明此表达产物具有特异的DNA结合活性。
The cDNA 1119 bp fragment (1105-2224) of the androgen receptor (AR), including its DNA binding domain to the partial hormone binding domain, was cloned into the expression plasmid pGEX, induced by IPTG, in E. coli. The GST-AR fusion protein was expressed in E. coli and partially purified by glutathione-Sepharose 4B affinity chromatography. A C3 (1) DNA fragment with activity of androgen responsive element (ARE) was used as a positive probe. The gel block analysis and DNaseI footprinting demonstrated that the expressed product had specific DNA binding activity.