β人绒毛膜促性腺激素在毕赤酵母中的表达及鉴定  

Identification of expressed human chorionic gonadotropin β subunit in pichia pastoris

作　　者：郑俊峰[1] 王桂玉[2] 于和鸣[2] 马旭[2] 

机构地区：[1]中国协和医科大学,北京100730 [2]国家人口计生委科学技术研究所,北京100081

出　　处：《生殖医学杂志》2005年第1期36-39,共4页Journal of Reproductive Medicine

摘　　要：目的　在毕赤酵母表达系统中获得β人绒毛膜促性腺激素(hCG)高效分泌表达,并鉴定其抗原活性。　方法　将βhCG基因亚克隆入 pPIC9K表达载体,线性化后转化 GS115 酵母细胞,经 G418 抗性筛选获多拷贝重组子并诱导表达。表达产物采用SDS 聚丙烯酰胺凝胶电泳(SDS- PAGE)、Western blot、酶链免疫吸附试验(ELISA)鉴定。　结果　经甲醇诱导,酵母细胞表达出βhCG,此蛋白在SDS PAGE电泳上显示为一条分子量为32×103 阳性条带,在Western blot试验中能被βhCG多抗识别,在竞争抑制性ELISA中能抑制βhCG单抗与 hCG全分子的结合。　结论　βhCG在毕赤酵母中得到高效表达,并形成正确的蛋白结构。Objective: To express human chorionic gonadotropin β subunit (β hCG) in pichia pastoris and study its immunological characteristics. Method: Subcloned β hCG gene was joined with pPIC9K yeast expression vector. The recombinant β hCG was induced expression by methanol in pichia pastoris and the product was identified by SDS-PAGE, Western blot and ELISA. Result: The β hCG was expressed in yeast cells and observed as a band of 32×103 with SDS-PAGE and western blot. Conclusion: The β hCG can be efficiently expressed in yeast and show a correct protein folding.

关 键 词：人绒毛膜促性腺激素 毕赤酵母 表达载体 

分 类 号：Q789[生物学—分子生物学]