Animal cell screening system based on GS efficient expression vector and its application
10.7644/j.issn.1674-9960.2014.10.015
- VernacularTitle:基于GS筛选系统的动物细胞高效表达载体的构建及应用
- Author:
Zhaogang GAO
;
Yong SHAO
;
Lihua GAO
;
Yun PAN
;
Yu LIU
;
Yipei LI
;
Xianwen HU
;
Huipeng CHEN
- Publication Type:Journal Article
- Keywords:
glutamine synthetase;
methotrexate;
methionine sulphoximine;
cytomegalovirus;
efficient expressing cell lines;
recombinant proteins
- From:
Military Medical Sciences
2014;(10):807-810
- CountryChina
- Language:Chinese
-
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.
