Construction Of Human Bone Morphogenetic Protein-2 Eukaryotic Expression Vector And Transfection Of Rabbit's Bone Mesenchymal Stem Cells

Objective:(1)To construct human bone morphogenetic protein-2(hBMP2) eukaryotic expression vector.(2)To dectect the expression of hBMP2 in rabbit's bone mesenchymal stem cells.Methods:(1)Human BMP2 gene was amplified by RT-PCR method from human osteosarcoma cells and constructed into eukaryotic expression vector pcDNA3.1- hBMP2.(2)Human BMP2 gene in the vector pcDNA3.1- hBMP2 was identified by PCR amplification,enzym edigestion and DNA sequencing.(2) The expression of human BMP2 gene was dectected by RT-PCR and immunohistochemical analysis after transfection of rabbit's bone mesenchymal stem cells.Result:(1)The cloned DNA was confirmed to be Human BMP-2 gene by PCR amplifcation,enzym edigestion and DNA sequencing.(2)The expression of human BMP2 gene was confirmed by RT-PCR and immunohistochemical analysis after transfection of rabbit's bone mesenchymal stem cells.Positive findings were not found in those cells that were not transfected.Conclusion:(1)In this study,hBMP2 gene eukaryotic expression vector was successfully constructed and can be expressed in bone mesenchymal stem cells after transfection.(2)This study provids the foundation of using BMP2 gene therapy to accelerate new bone formation in distraction osteogenesis.