军事医学Issue(10)：807-810,4.DOI:10.7644/j.issn.1674-9960.2014.10.015
基于GS筛选系统的动物细胞高效表达载体的构建及应用
Animal cell screening system based on GS efficient expression vector and its application
摘要
Abstract
Objective To obtain highly expressing cell lines by inserting the glutamine synthetase (GS) screening system and replacing the promoter of the vector.Methods The mutation of the point BamHⅠwas induced to build a new vector pIRES2-EGFP.The marker gene GS was inserted by AseⅠ and NheⅠ, and the promoter hCMV was replaced by PacⅠand NheⅠ.The new vector pHGS1.0 and the vector pIRES2-enhanced screen fluorescein protein( EGFP)-B were inserted by the recombinant protein TEM8 ( 1-227 )-VEGFR1 domain2-IgG2 ( TV-IgG2 ) gene to analyze the advantages of the expression.Results The glutamine synsthetase is successfully inserted, the human cytomegalovirus replaced, and recombinant protein is increased 5-fold by human immunoglobulin quantification kit.Conclusion The GS system is a highly protein expressing system.关键词
谷氨酰胺合成酶/甲氨喋呤/蛋氨酸亚氨基代砜/巨细胞病毒/高效表达细胞系/重组蛋白质类Key words
glutamine synthetase/methotrexate/methionine sulphoximine/cytomegalovirus/efficient expressing cell lines/recombinant proteins分类
生物科学引用本文复制引用
高招刚,邵勇,高丽华,潘芸,刘羽,李伊培,胡显文,陈惠鹏..基于GS筛选系统的动物细胞高效表达载体的构建及应用[J].军事医学,2014,(10):807-810,4.基金项目
“重大新药创制”国家科技重大专项资助项目 （）
