BMPs Promote Bone Differentiation And The Related Mechanism

PART ONE CONSTRUCT THE RECOMBINANT ADENOVIRUS OF Ad5-BMP9Objective : Construct the recombinant adenovirus plasmid Ad5-BMP9 expressing BMP9 , then transfect the plasmid into the package cell HEK293 and generate the recombinant adenovirus .Methods: Firstly, we amplify the target gene BMP9 containing the linker through the PCR , then intergrate the target gene into the shuttle vector of pAdTrack-CMV by restrict ligation . Secondly , recombine the plasmid in the competent E coli containing the adenovirus skeleton plasmid Adeasy -1 to get Ad5-BMP9 plasmid . Finally , we transfect the recombinant plasmid Ad5-BMP9 into the package cell HEK293 and generate the high titre adenovirus .Results : Construct the adenovirus plasmid Ad5-BMP9 successfully , generate the adenovirus expressiong BMP9 and the titre was about 1.5×1011pfu/ml.Conclusion : Recombining of the plasmid in the E coli cells is an effective method for construct the adenovirus plsmid . We cloned the...