Construction of hTNFα/CHO genetic engineered cell and verification of its protein secretion function
10.11659/jjssx.01E016024
- VernacularTitle:hTNFα／CHO基因工程细胞的构建及其分泌蛋白功能的检定
- Author:
Lei TIAN
;
Jingkun PAN
;
Ming SHI
;
Yilong XUE
- Publication Type:Journal Article
- Keywords:
genetic engineered cell;
human tumor necrosis factor;
transfection;
protein secretion
- From:
Journal of Regional Anatomy and Operative Surgery
2016;25(4):240-243
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the genetic engineered cell line which can continuously secrete human tumor necrosis factor ( hTNFα) in Chinese Hamster Ovary cells( CHO cells) ,and observe the change of its protein secretion function.Methods Constructed plas-mid which carries hTNFαgene expression through vector GV141.Selected stable transfection cell lines by G418 and transfection with lipo-fectamine.Identified its gene expression with Real-Time PCR,and identified its protein secretion by ELISA.Results GV141-hTNFαexpres-sion vectors were constructed successfully which were proved by sequence alignment.Real-Time PCR proved that it contained hTNFαgene in hTNFα/CHO cell line.ELIAS identification results showed that the cell lines can continuously secrete hTNFαwithin a certain cell propaga-tion.Conclusion The hTNFα/CHO cell line can continuously secrete human tumor necrosis factor within a certain cell propagation.
