Construction of pEGFP-VEGF recombinant plasmid and its expression in osteoblast
- VernacularTitle:pEGFP-VEGF重组质粒的构建及其在成骨细胞中的表达
- Author:
Wei FAN
- Publication Type:Journal Article
- Keywords:
VEGF;
Plasmid construction;
Gene transfection;
Osteoblast
- From:
Journal of Chongqing Medical University
1987;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct a vector containing human vascular endothelial growth factor,VEGF 121 (VEGF 121 ) with green fluorescence protein as the reporter gene,and then to observe its expression in osteoblast.Methods:VEGF 121 gene was amplified from foetus brain library by PCR,then ligated with pEGFP-C1 to form recombinant plasmid pEGFP-VEGF and transfected into osteoblast by liposome.The green fluorescence protein expression was observed by fluorescence microscopy and the VEGF expression was last detected by immunocytochemistry and ELISA.Results:VEGF 121 gene fragment was amplified by PCR and ligated to pEGFP-C1,then transfected into osteoblast successfully.The strong expression of green fluorecensce protein was observed by fluorecensce microscopy,and the expression of VEGF protein was detected by immunocytochemistry and ELISA.Conclusion:Recombinant plasmid pEGFP-VEGF is effectivly expressed after being transfected into osteoblast in vitro,which provides theoretical support for further study with its simple and reliable detecting methods.
