Recombinant human bone morphogenetic protein-7 gene adeno-associated virus vector and its biological activity in vitro.docVIP

 PAGE \* MERGEFORMAT 18 Recombinant human bone morphogenetic protein-7 gene adeno-associated virus vector and its biological activity in vitro OF: When Zhibin, strong Hui, Fan Lihong, Kun-Zheng, Xiao Qian Party [Abstract] Objective To construct green fluorescent protein (GFP) tag and carrying human bone morphogenetic protein-7 (hBMP 7) gene in non-pathogenic recombinant adeno-associated virus (AAV), as a gene carrier for its bone marrow stem cells infected with (BMSCs), detection of the virus in vitro biological activity and function. methods with hBMP 7 gene from pBluescript KS hBMP7 hBMP 7 plasmid was amplified fragments, recombinant backbone plasmid pAAV hBMP7 IRES GFP. this plasmid and the AAV packaging plasmid pAAV RC , helper plasmid pAAV Helper AAV 293 cells were transfected by homologous recombination adeno-associated virus rAAV hBMP7 IRES GFP. infected with AAV HT1080 virus titer, the application of the recombinant virus in vitro cultured rabbit BMSCs, fluorescent cell counting assay Best virus multiplicity of infection, Western blot assay gene expression in vitro BMP 7, osteogenic assay in vitro biological activity of BMP 7. Results Expression of recombinant adeno-associated virus plasmid by enzyme digestion, sequencing is correct. Recombinant adeno-associated virus titer of approximately 3.6 1011v.p./mL. recombinant virus multiplicity of infection of rabbit BMSCs was the best 5 104 vp / cell. Western blot showed that recombinant virus rAAV hBMP7 IRES GFP cells transfected with BMP expression in BMSCs . osteogenic rAAV hBMP7 IRES GFP experiments confirmed the expression of BMP in vitro activity with good bone. Conclusions The preparation of the rAAV hBMP7 IRES GFP viruses, experiments with high titers of viral vectors, infection, and good bone tissue engineering to meet need for further studies in vivo and in vitro gene therapy provide a favorable environment for bone healing tool. [Keywords:] human bone morphogenetic protein 7; adeno-assoc