Construction Of Recombinant Plasmid PEGFP-N1-apoJ And Its Expression In Rat Bone Marrow Mesenchymal Stem Cells In Vitro

Objective: To construct recombinant plasmid pEGFP-N1-apo J,transfect rat bone marrow mesenchymal stem cells,and detect its expression in rat bone marrow mesenchymal stem cells(MSCs)in vitro.Methods: Rat bone marrow MSCs were transiently transfected with the recombinant plasmid pEGFP-N1-apoJ and transfection efficiency of the Green Fluorescent Protein(GFP)was determined.Real-time PCR ?Western Blot and immunocytochemistry analysis were also performed to detect the expression of apoJ in rat bone marrow MSCs.Results: Transient transfection efficiency can reach 35%.After transfection,transcription of apoJ was detected in MSCs and expression of apoJ protein was also verified.Conclusion: Recombinant plasmid pEGFP-N1-apo J was successfully constructed,transfected into rat bone marrow MSCs and expressed in rat bone marrow MSCs.This procedure may be helpful to the application of apoJ for gene therapy and provide the experimental basis for the further study of apoJ as a new treatment of cerebral hemorrhage.