Cloning of human vascular endothelial growth factor cDNA and its expression in rabbit osteoblasts
- VernacularTitle:人血管内皮细胞生长因子的cDNA克隆及其在兔成骨细胞中的表达
- Author:
Shuhua YANG
;
Cao YANG
;
Weihua XU
- Publication Type:Journal Article
- Keywords:
Endothelial growth factors;
Blood vessels;
Gene amplification;
Gene expression
- From:
Chinese Journal of Orthopaedics
2000;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone VEGF gene and construct its eukaryotic expression vector for the evaluation of the possability of VEGF gene therapy in orthopedic disease. Methods Human vascular endothelial growth factor(hVEGF) cDNA was amplified by nested PCR method from the HL60 cells and cloned to expression vector pcDNA 3. The cDNA was identified by enzyme digestion and DNA sequencing. Rabbit osteoblasts were transfered with pCD-hVEGF165 plasmid by lipofectin mediated gene transfer method. The transient expression of VEGF were detected by Streptavidin-Biotin-enzyme Complex(SABC). Results The cloned cDNA was confirmed to be VEGF165 cDNA. It was observed that the expression of human VEGF gene was detected distinctly 72 h after transfering. Conclusion We successfully cloned hVEGF 165 gene and construced its eukaryotic expression vector, which provided the further foundation of VEGF gene therapy for ostenecrosis,bone defeat and fracture.
