Construction of eukaryotic expression carrier of recombinant pcDNA3.1-hBMP-7 and transfection into bone marrow stromal cells of rabbit

[Objective]To construct arecombinant eukaryotic expression plasmid carrying human BMP-7 gene which was transfected into bone marrow stromal cells(BMSCs) in vitro and expressed.[Method]Clone human BMP-7 cDNA from a Chinese woman placenta was recombinanted with plasmid pcDNA3.1,to construct eukaryotic expression carrier of recombinaut pcDNA3.1-hBMP-7.The BMSCs were isolated from rabbit bone marrow and cultured in vitro.They were divided into three groups: pcDNA3.1-hBMP-7 transduced group;pcDNA3.1 transduced group;untransduced group.The expression of hBMP-7 was detected by RT-PCR,Immuneohistochemistry.ALP,Collogen,Osteocalcin production were detected.[Result]The pcDNA3.1-hBMP-7 transduced BMSCs expressed hBMP-7 at both mRNA and protein levels.ALP activity was detected in pcDNA3.1-hBMP-7 transduced cells from day 2 to day 10,peaking on day 8.Collogen,osteocalcin production were elevated significantly too(P0.05).[Conclusion]Eukaryotic expression carrier of recombinant pcDNA3.1-BMP-7 was constructed successfully.The results indicate that hBMP-7 is expressed in BMSCs sufficiently and is involved in inducing differentiation of BMSCs into osteoblast.The method would provide substantial basement for hBMP-7 gene therapy.