CLONNING OF HUMAN GM-CSF GENE AND ESTABLISHMENT OF ITS STABLE EXPRESSION CELL LINE

ObjectiveEstablish a stable cell line to produce human granulocyte-macrophage colony-stimulating factor(hGM-CSF) with high level expression in order to reduce production cost.MethodsAmplify GM-CSF cDNA with RT-PCR method;Transfect gene into L1/2 cell line by electroporation;Identify gene products by Western Blotting,ELISA and biological assay.ResultsHuman GM-CSF cDNA expression vector was prepared by inserting the hGM-CSF cDNA into pcDNA3.1A.Stable cell line expressing human GM-CSF was established by transfecting L1/2 cells with electroporation,following by G418 selection.The gene product was identified to have biological activity.The average yield of hGM-CSF was 850 ng/10~6 cell.ConclusionThe established cell line can highly-productive express of hGM-CSF with biological activity.