Co-expression of BMP-2 and TGF-β1 genes in rabbit marrow stromal cells

Objective To investigate co0expression of BMP-2 and TGF-β1 genes of the bicistronic eukaryotie expression plasmid plRES-BMP-2-TGF-β1 in rabbit marrow stromal cells(MSCs).Methods Using PCR technique,the DNA fragments containing two new enzyme digestion sites of BMP-2(1 188 bp) and TGF-β1(1 173 bp)genes were obtmned from pGEM-T-BMP-2 and pGEM-T-TGF-β1 plasmids.then inserted into bieistronic eukaryotie expression plasmid pIRES.The inserted target genes in the plasmid were verified by restriction enzyme digestion and nucleotide sequencing analysis.MSCs were transfected with this bicistronic eukaryotie expression plasmid using hpofeearmin reagent.The expression of BMP-2 and TGF-β1 were detected by RT-PCR.Results The directions and sequences of BMP-2(1188 bp)and TGF-β1(1173 bp)genes in plasmid pIRES-BMP-2-TGF-β1 were correct and BMP-2 and TGF-β1 mRNAs could be detected in MSCs.Conclusion BMP-2 and TGF-β1 genes of the bicistronie eukaryotic expression plasmid plRES-BMP-2-TGF-β1 Can be co-expressed successfully in MSCs.which can be used for further investigation of co-gene therapy of articular cartilage iniury. Key words: BMP-2; TGF-β1;  Marrow stromal cells: Gene expression