Expression of human IL-10 cDNA in E. coil

Objective To clone the full length of human interleukin-10 cDNA,construct prime expression vector pMon-IL10 and induce its expression in E. coli.Methods Total RNA was extracted from ConA-stimulated human peripheral blood mononuclear cells (PBMC). The cDNA of human IL-10 was cloned from RNA by RT-PCR and subcloned into pMon vectors. The E. coli DH5α cells were transfected with the constructed pMon-IL10 recombinant, and the conbinant human IL-10 expression in E. coli was induced by nalidixic acid. Finally, the expressed IL-10 protein was assayed by SDS-PAGE and Western blot.Results The transcription of Il-10 in PBMC was stimulated by ConA, making the cloning of IL-10 cDNA easier. Gene sequencing confirmed that the sequence of human IL-10 cDNA cloned was identical to that reported in Genbank. Under nalidixic acid induction, the recombinant human IL-10 was effectively expressed in E. coli, and the expressed protein could specifically combine with the rabbit anti-human IL-10 antibody.Conclusion The full-length of IL-10 cDNA has been successfully cloned and expressed in E. coli DH5α.