Cloning, expression and identification of human interleukinª²10 gene

    SUN Jie, LI Boª²an, QI Yang, ZHOU Jiª²wen, ZHU Lei, CHI Shuª²ping, CHENG Yun*

    Department of Immunology, Beijing Institute of Infections Diseases, Beijing 100039, China

    [Abstract] AIM: To construct a prokaryotic expression vector containing human interleukinª²10 (ILª²10) gene, express and identify the protein. METHODS: Human ILª²10 gene from HepG2 was isolated and identified by DNA sequencing, and then cloned into the expression vector PETª²28 a (+) to construct prokaryotic expression vector. The recombinant protein was expressed in BL21(DE3) and identified by Western blot and ELISA. RESULTS: The expressed protein was mainly located in the inclusion body. The relative molecular mass of the expressed product was identical to that of prediction. The expressed protein had binding activity with specific antibody. CONCLUSION: The human ILª²10 gene was successfully cloned and expressed in E.coli. The expressed product had antigenicity which provides foundation for preparation of monoclonal antibodies against ILª²10. ......