Expression of Eukaryotic Vector Carrying TGF-β1 and EGFP Genes in BMSCs.

Objective:To construct a bicistronic eukaryotic co-expression plasmid carrying TGF-β1and enhanced green fluorescent protein(EGFP) and observe its expression in bone mesenchymal stem cells(BMSCs)in vitro.Methods: TGF-β1 expression unit was inserted into pIRES2-EGFP to produce the bicistronic eukaryotic co-expression plasmid.The recombinant plasmid and pIRES2-EGFP were transfected into BMSCs,respectively,which were detected by fluorescence microscope and cell immunohistochemical staining.The untransfected BMSCs served as control group.Results: The eukaryotic vector carrying TGF-β1 and EGFP genes was successfully constructed.Positive clones were obtained by G418 screening after 10-15 days.The clones group which was transfected by pTGF-β1-IRES2-EGFP expressed EGFP and TGF-β1 simultaneiously.However,some clones(20%) only expressed TGF-β1 but no EGFP.Another group which was transfected by pIRES2-EGFP expressed EGFP but no TGF-β1.Conclusion: The bicistronic eukaryoutic vector containing TGF-β1 and EGFP could express TGF-β1 aod EGFP in some of BMSCs,which provided a method to study the migration and the osteogenesis of TGF-β1 gene-modified BMSCs in vivo.