Expression and significance of the recombinant rat GATA4-pEGFP gene in bone marrow mesenchymal stem cells

Objective To screen bone marrow mesenchymal stem cells(MSCs) expressing GATA4-pEGFP in vitro,so as to provide theoretical foundation for the research of myocardial infarction.Methods Total RNA was extracted from embryonic myocardial cells,GATA4 gene was amplified by RT-PCR,the target gene was cloned to pEGFP-N3 vector to get recombinant GATA4-pEGFP plasmid,the bone marrow mononuclear cells was derived and MSCs were obtained by gradient centrifugation.The recombinant plasmid was transfected into MSCs using lipofectamine 2000,then screened for 2 weeks by G418,Western blot and immunofluroscence staining were used to examine the expression pattern of transfected protein.Results Recombinant rat GATA4-Pegfp eukaryocyte expression vector was constructed and transfected into MSCs successfully,Western blot and immunofluroscence staining showed that GATA4-Pegfp gene expressed positively in the MSCs.Conclusion MSCs expressing GATA4-pEGFP can be constructed successfully,this provides opportunity to study and treat myocardial infarction using tissue engineering technologies.