The Research Of Constructhuman Bone Morphogenetic Proteins Factor-2 Eukaryotic Plasmid Expression In Marrowstromal Cells

Objective After constructing the eukaryotic expression plasmid of hBMP-2, we transfered it into rabbit marrow stromal cells to make a stable expression, so that we could investigate the function of human bone morphogenetic proteins factor-2 (hBMP-2) in distraction osteogenesis (DO), especially in the bone tissue engineering.Methods 1.The hBMP-2 cDNA was amplified and cloned into the eukaryotic plasmid pcDNA3.1 and screened with Escherichia coli JM109.2.The recombinant plasmid was transfered into the rabbit marrow stromal cells by lipofectamine, and the positive cell clones were selected with G418.Results 1.The new recombinant plasmid was digested with EcoRI, and the electrophoresis of the digested products showed two fragments:1.5kb fragment and 5.5kb. 2.It was showed by the RT-PCR method that the hBMP-2 gene transfected MSCs prominently elevated mRNA expression of hBMP-2.It was confirmed that hBMP-2 gene could be expressed effectively in MSCs transfected with the recombinant plasmid.Conclusion The pcDNA3.1- hBMP-2, a eukaryotic expression plasmid of hBMP-2 gene has been constructed. The hBMP-2 gene could be expressed effectively in rabbit marrow stromal cells transfected with the recombinant plasmid.