Study on cloning and eukaryotic expression of human interleukin 10 gene

  Objective   To construct a eukary otic expression vector containingh IL-10 cDNA and express it in CHO cells.

  Methods   hIL-10 cDNA, which was amplified by RT-PCR from human peripheral blood mononuclear cells, was cloned to vector pMD18-T.After the sequence was confirmed, the hIL-10 cDNA was isolated and then was inserted into eukaryotic expression vector pCIneo.The recombinant plasmid pCIneo-hIL-10 was transfected into CHO cells with Lipofectamine Plus^TM Reagent.The expression of hIL-10 in CHO cells was tested by RT-PCR and immunohistochemistry assay; the secretion of hIL-10 in medium was tested by ELASA and western blot assay.

  Results   The sequence of hIL-10 cloned was identical with that was published on GenBank.Mammalian recombinant expression plasmid pCIneo-hIL-10 was constructed.And the transcription and expression of hIL-10 were detected.

  Conclusion   The eukaryotic expression system of hIL-10 was successfully established, which might provide the basis for studying affection of IL-10 in clinics and production of gene engineering medicine.