Expression of Human Connexin26 in COS-7 Cells
- VernacularTitle:人连接蛋白Connexin26基因的克隆、表达及初步鉴定
- Author:
Cong XU
;
Guowei LI
;
Zeyu LIU
;
Jiawei LIAO
;
Panpan XUE
;
Binyan TAN
;
Jiahai LU
- Publication Type:Journal Article
- Keywords:
genetic deafness;
Connexin26;
eukaryotic expression vector;
gene therapy
- From:
Journal of Tropical Medicine
2007;7(5):401-407
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the eukaryotic expression plasmid for the expression of human Connexin26 in COS-7 cells.Methods Total RNA was isolated from human peripheral blood lymphocytes and used as template for the PCR cloning of the human Connexin26 gene.The human Cx26 cDNA containing the 678 bp whole coding region of the human Connexin26 gene was amplified by PCR using specific primers and cloned into the pCI-neo vector to construct the recombinant eukaryotic expression plasmid,pCI-Cx26.The recombinant plasmid was identified by restriction endonuclease digestion,and transfected into COS-7 cells by liposome.The expression of Cx26 mRNA and the protein were analyzed by RT-PCR and SDS-PAGE,respectively.Results Restriction endonuclease digestion analysis verified successful construction of the recombinant plasmid,pCI-Cx26.The expression of Cx26 mRNA and protein in the transfected COS-7 cells were detected by RT-PCR and SDS-PAGE,respectively.Conclusion The eukaryotic expression plasmid for human Cx26 has been constructed successfully with the capability of expression in COS-7 cells.
