表达血小板膜糖蛋白GPIb-Ⅸ复合物的中国仓鼠卵巢细胞株研究模型的建立  

Establishment of Chinese Hamster Ovary Cell Line Expressing Recombinant GPIb-Ⅸ Complex

作　　者：廖翊[1] 张卫林[1] 元艳宏[1] 史权威[1] 李素萍[1] 闫荣[1] 王志成[1] 戴克胜[1] 

机构地区：[1]北京航空航天大学生物与医学工程学院,北京100191

出　　处：《中国实验血液学杂志》2009年第6期1560-1563,共4页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目,编号:30770795

摘　　要：本研究旨在建立表达重组野生型和突变型血小板膜糖蛋白GPIb-Ⅸ复合物的中国仓鼠卵巢(Chinese Hamster Ovary,CHO)细胞研究模型,为进行GPIb-Ⅸ生理功能的相关研究奠定基础。从表达野生型或缺失突变型GPIbα的大肠杆菌宿主菌中抽提质粒并用EcoRI酶切鉴定,然后将分别表达GPIbα、GPIbβ和GPIX3种基因的质粒共转染到中国仓鼠卵巢细胞中,通过免疫共沉淀、Western blot和流式细胞仪检测其表达效果。结果表明,转染48小时后,CHO细胞裂解物和细胞表面均可检测到GPIb-Ⅸ复合物的表达。结论:成功构建了表达血小板膜糖蛋白GPIb-Ⅸ复合物的CHO细胞研究模型。The aim of this study was to construct Chinese Hamster Ovary （CHO） cell models expressing recombinant wild-type GPIb-IX and mutant GPIb-IX complex, so as to provide the platform to study the related physiologic functions of GPIb-IX. The plasmids were extracted from E. coli expressing wild-type or deletion mutant GPIbα and were identified by digestion with EcoR I. Three plasmids containing GPIbα, GPIbβ, and GPIX genes were co-transfected into CHO cells, and then the expression of GPIb-IX complex was detected by immune coprecipitation, Western blot and flow cytometry. The results showed that the expression of GPIb-IX complex could be detected in the lysate and on the surface of CHO cells at 48 hours after transfection. In conclusion, CHO cell models expressing recombinant wild-type or mutation GPIb-IX complex has been successfully constructed.

关 键 词：血小板膜糖蛋白 GPIb—IX复合物 CHO细胞株 

分 类 号：R331.143[医药卫生—人体生理学]