Construction of eukaryotic expression vectors of nephroblastoma overexpression gene and expression in COS-7 cells
- VernacularTitle:NOV基因真核表达载体的构建及在COS-7细胞中的表达
- Author:
Chengren LI
;
Wenqin CAI
;
Bingyin SU
;
Chenggang ZHANG
- Publication Type:Journal Article
- Keywords:
nephroblastoma overepression gene;
RT-PCR;
gene cloning
- From:Journal of Third Military Medical University
2002;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To obtain eukaryotic expression vectors containing coding region of nephroblastoma overexpression gene (NOV) and detect its expression in COS-7 cells. Methods A 1 165-bp cDNA fragment was amplified from the total RNA of normal rat brain tissue by RT-PCR and cloned into eukaryotic expression vector pcDNA3.1/Myc-His(+)/lacZ. The cloned insert was identified by double digestion of the recombinant plasmid with restriction enzymes HindⅢ and BamHⅠ. The recombinant plasmid was transfected into COS-7 cells with liposome. The expression of NOV gene was detected by Western blotting and immunocytochemistry. Results Eukaryotic expression vectors containing 1 165 -bp coding region of NOV gene was constructed. COS-7 cells transfected with the recombinant plasmid expressed high level of NOV protein in cytoplasm. Conclusion That eukaryotic expression vectors containing coding region of NOV gene was constructed can provide a strong molecular tool for the studies of effect of NOV gene.
