关键词: 白细胞介素2, 基因克隆, 原核表达

Abstract: Abstract:Objective To establish E.coli strain expressing human IL-2 and MUC1 fusion protein to provide experimental fundation for a novel MUC1 cancer vaccine.Methods Human IL-2 gene fragments were obtained by PCR and they were inserted into recombinant vector pBS-SK-MUC1.After digested,human IL2-MUC1 gene fragments were cloned into prokaryotic expression vector pGEX-4T-1 and E.coli BL21 was transformed by constructing pGEX-IL2-MUC1.IL2-MUC1 fusion protein expression was induced by IPTG and analyzed by SDS-PAGE and Western blotting.Results Acquired gene fragments of human IL-2 (402 bp) and tandem IL2-MUC1 (852 bp)　 were identified by digestion and DNA sequencing. The recombinant vector of pGEX-IL2-MUC1 was constructed successfully.SDS-PAGE result proved that GST-IL2-MUC1 fusion protein with a relative molecular mass of about 64 000 was expressed.Western blotting result showed that the expressed products had specific reaction with both rabbit anti-human MUC1 polyclonal antibody and mouse anti-human IL-2 monoclonal antibody.Conclusion The engineering E.coli strain expressing human IL2-MUC1 fusion protein is successfully established.It is a basis to further study on novel cancer vaccine with MUC1 as target.

Key words: interleukin 2, gene cloning, prokaryotic expression