Cloning & expression of human IL-24 in COS-7 cells

Objective:To clone human IL-24 gene,and construct its eukaryotic expression vector,and express it in COS-7 cells detect its anti-tumor activities of recombinant hIL-24 protein.Methods:The total RNA was extracted from peripheral blood mononuclear cells.And the functional fragment of human IL-24 gene was amplified by RT-PCR.It was cloned into pUC19 sequenced and was subcloned into pcDNA3 vector.Constructed pcDNA3-hIL-24 was identified by endonucleases digestion PCR.The recombinant expression plasmids was transfected into COS-7 cells,human hIL-24 expressed in COS-7 cells was deteced by RT-PCR.The apoptosis-inducing activities of recombinant protein hIL-24 was tested by MTT assey,TUNEL assey FCM assey.Results:Obtained full encoding sequence of hIL-24 was identical with that included in Genbank, the eukaryotic expression vector pcDNA3-hIL-24 was constructed correctly.Expression of human IL-24 in COS-7 cells was identified by RT-PCR.The apoptosis of A549 cells induced by hIL-24 was proved by TUNEL FCM assay.Conclusion:The successful cloning,transient expression preliminary study of apoptosis effect of human IL-24 protein provide experimental basis for the further studying of molecular mechanism and clinical application of hIL-24 on anti-tumors.