人源雄激素受体DNA结合结构域的原核表达与其响应元件的复合物结晶  

Prokaryotic expression of homo androgen receptor DNA binding domain and co-crystallization with androgen response element

作　　者：秦童 黄震 QIN Tong;HUANG Zhen(Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education,College of Life Sciences,Sichuan University,Chengdu 610065,China)

机构地区：[1]四川大学生命科学学院生物资源与生态环境教育部重点实验室

出　　处：《四川大学学报（自然科学版）》2019年第6期1151-1156,共6页Journal of Sichuan University(Natural Science Edition)

基　　金：国家自然科学基金(21572146)

摘　　要：为获得高分辨率的雄激素受体与其响应元件的复合物晶体结构,为前列腺癌的治疗提供药物靶点设计的理论基础.本研究构建了AR-DBD的原核表达载体,在0.5 mmol/L IPTG,16℃条件下诱导24 h表达该蛋白.通过与AR响应元件寡核酸的结合优化蛋白纯化条件,使AR-DBD蛋白易于纯化.最终,蛋白核酸复合物晶体呈现出边缘清晰,短棒状的立体结构,本研究制备的蛋白核酸复合物晶体可直接用于X射线衍射分析.Obtained a high-resolution complex crystal structure of the androgen receptor(AR)and its response element(ARE)could provide a theoretical basis for drug target design for the treatment of prostate cancer.In this study,a prokaryotic expression vector of AR-DBD was constructed,and the protein was inducted to expression at 16℃ with 0.5 mmol/L IPTG for 24 h.In order to purify AR-DBD protein easily,the protein purification conditions were optimized by binding to ARE oligo-nucleic acids.Finally,we obtained the protein-nucleic acid complex crystal that exhibited short rod-like three-dimensional structures with clear crystal edges.The protein-nucleotide complex crystals prepared in this study can be used for X-ray diffraction analysis directly.

关 键 词：雄激素受体 前列腺癌 DNA结合结构域 蛋白核酸复合物晶体 原核表达 

分 类 号：Q78[生物学—分子生物学]