【摘要】 目的:在巴氏毕赤酵母细胞中表达hCGβ-(CTP)n(n=3,4)-3C3d融合蛋白。方法:将(CTP)n(n=3,4)基因与分子佐剂C3d基因的三聚体连接后,克隆入载体pPIC9K得到表达质粒pPIC9Khis6(CTP)n(n=3,4)3C3d,经限制性内切酶酶切鉴定及DNA测序确认后电转入酵母细胞中进行表达。结果:在酵母细胞内检测到目的基因的表达产物,Westernblot结果显示它们的分子量分别为125kD和138kD左右。结论:运用巴氏毕赤酵母表达系统成功表达了hCGβ-(CTP)n(n=3,4)-3C3d融合蛋白,为进一步研究其免疫原性和用作抗肿瘤疫苗奠定了基础。更多还原

【Abstract】 Objective: To investigate the expression of the gene enconding fusion protein hCGβ-(CTP)n (n=3,4)-3C3d in the methylotropic yeast Pichia pastoris. Methods: We constructed the expression vectors pPIC9Khis6 (CTP)n (n=3,4)3C3d, using the recombinant genes of (CTP)n (n=3,4) and triplont of molecular adjuvant 3C3d. After identification by restriction endonuclease digestion and DNA sequencing, the expression vectors were transferred into GS115 cells by electroporation. Results: Western blot results suggested that the expressed proteins with molecular weights of about 125 kD and 138 kD each existed in the GS115 cells. Conclusion: The fusion protein hCGβ-(CTP)n(n=3,4)-3C3d was successfully expressed in Pichia pastoris. This study prepared basic necessary data for further investigation of the immunogenicity and anti-cancer vaccine of fusion protein hCGβ-(CTP)n (n=3,4)-3C3d.更多还原