Construction of eukaryotic expression vector of rat EPO and its expression in bone marrow mesenchymal stem cells / 第三军医大学学报

ABSTRACT

Objective To construct eukaryotic expression plasmid pEGFP-N1/EPO and to detect its expression in vitro in order to provide experimental evidence for further study on the treatment of hypoxic-ischemic brain damage.Methods The total RNA was extracted from rat kidney.EPO cDNA fragment was obtained by RT-PCR amplication,inserted into pEGFP-N1 vector and identified by restriction endonuclease digestion and nucleotide sequencing.MSCs cells were transfected with the plasmid using Lipofectamine 2000.The expression of EPO protein was detected by immunohistochemistry technique and RT-PCR.Results Confirmed by restriction endonuclease digestion and sequencing,the recombinant plasmid contained the sequence of rat EPO gene was successfully constructed.After transfection with the plasmid,EPO protein could be expressed in MSCs cells.Conclusion The constructed eukaryotic expression vector pEGFP-N1/EPO can express rat EPO in vitro.