[Cloning, expression and identification of human interleukin-10 gene].

AIM

To construct a prokaryotic expression vector containing human interleukin-10 (IL-10) gene, express and identify the protein.

METHODS

Human IL-10 gene from HepG2 was isolated and identified by DNA sequencing, and then cloned into the expression vector PET-28 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 IL-10 gene was successfully cloned and expressed in E.coli. The expressed product had antigenicity which provides foundation for preparation of monoclonal antibodies against IL-10.