生物技术通报2016，Vol.32Issue(5)：146-150,5.DOI:10.13560/j.cnki.biotech.bull.1985.2016.05.019
磷脂酰肌醇蛋白聚糖3真核表达载体的构建及在细胞中的表达
Construction of Eukaryotic Expression Vector for Glypican-3 and Its Expression in Cell
摘要
Abstract
This work aims to construct the eukaryotic expression vector of glypican-3(GPC3)for acquiring anti-GPC3 monoclonal antibody. The GPC3 gene was amplified by PCR and was cloned into an expression vector p3XFLAG-CMV-14,and the expression vector pCMV-gpc3 was constructed. HEK293 cells were transfected with the vector by liposome,and the final results were detected by Western blot. The stably-expressed cells were collected and ground,and the high-purity GPC3 protein was obtained by affinity column. The eukaryotic expression vector of pCMV-gpc3 was constructed successfully. After the transfection to HEK293 cells,the monoclonal cell line of stably-expressed was gained by G418 screening. Western blot analysis revealed that the target protein expressed markedly. A large number of GPC3 proteins were obtained via genetic engineering and purifying technology,which laid a foundation for the biological function and application of the protein as well as the preparation of monoclonal antibody.关键词
磷脂酰肌醇蛋白聚糖3/真核表达/脂质体转染/亲和纯化Key words
glypican-3/eukaryotic expression/liposome transfection/affinity purification引用本文复制引用
程华,方向东,崔硕,吴萌,张昭军,李泽夏..磷脂酰肌醇蛋白聚糖3真核表达载体的构建及在细胞中的表达[J].生物技术通报,2016,32(5):146-150,5.基金项目
河北省科学院高层次人才资助项目（20150503LR62-9），河北省科学院科技计划项目 （）
