Expression,purification and activity identification of hTPO in Escherichia coli

Objective:To express and purify human thrombopoietin in Escherichia coli.Methods:The TPO cDNA coding was amplified from the human fetal liver cell by RT-PCR,the target cDNA was inserted into the polycloning region of pQE30expression vector,then transferred into the E.coli M15.The expression products was injected into the mice with thrombocytopenia and the change of the platelet' level in peripheral blood was detected.Results:SDS-PAGE analysis showed that the rhTPO protein was ex-pressed especially in E.coli M15harboring pQE30-TPO recombinant plasmid induced by IPTG,and the target protein was purified with histidine-tail through Ni-NTA affinity gel.The rhTPO was effective in treating carboplatin-induced thrombocytopenia in mice.Conclusion:The recombinant human thrombopoietin was successfully expressed in Escherichia coli.