The cloning of human interleukin-10 gene and its expression in eukaryotic cells / 医学研究生学报

ABSTRACT

Objective: To clone human interleukin 10(hIL-10) gene full-length cDNA sequence and to construct its eukaryotic expression vector in Chinese hamster ovary(CHO) cells. Methods: cDNA fragment encoding hIL-10 gene was amplified from human peripheral normal white blood cells by RT-PCR and confirmed by DNA sequencing.The ds-DNA was inserted into pcDNA3 vector.This recombinant expressing vector was transfected into CHO cell.The IL-10 molecules expressed were detected by ELISA and the role of inhibition of IL-10 was determined with MTT. Results: The cDNA encoding hIL-10 was cloned by RT-PCR and inserted into T-easy vector and sequence analysis verified that the cloned fragment was hIL-10 cDNA.The IL-10 was expressed in the CHO and it blocked the lymphocyte transformation.Conclusion: The eukaryotic expression plasmid was constructed successfully,which will contribute to further studies on the role of hIL-10 in autoimmunity,transplantation immunity and inflammatory disease.