Construction of human interleukin 10 eukaryotic expression vector and the expression

Objective To construct an eukaryotic expression vector containing hIL 10 cDNA and express it in HeLa cells.Methods hIL 10 cDNA amplified by RT PCR from human peripheral blood mononuclear cells was cloned to vector pMD18 T. After confirming the sequence, the hIL 10 cDNA was isolated and inserted into eukaryotic expression vector pCIneo. The recombinant plasmid pCIneo hIL 10 was transfected into HeLa cells with Lipofectamine Plus TM Reagent.Then the expression of hIL 10 in HeLa cells was tested by RT PCR and immunohistochemistry assay; the secretion of hIL 10 in medium was detected by ELISA and western blot assay.Results The sequence of cloned hIL 10 was identical with that published on GenBank. Mammalian recombinant expression plasmid pCIneo hIL 10 was constructed. And the transcription and expression of hIL 10 were detected.Conclusions We successfully established the eukaryotic expression system of hIL 10,which might provide the basic method for applying IL 10 in clinics and in producing gene engineering medications.