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.