hCGb-CTP多聚体-3C3d融合蛋白在巴氏毕赤酵母中的表达  

Expression of Fusion Protein hCGβ-CTP Polymer and 3C3d in Pichia pastoris Cells

作　　者：冯文华[1] 石燕[1] 喻光荣[1] 沈卫英[2] 蒋俶[2] 申庆祥[2] 

机构地区：[1]复旦大学上海医学院,上海200032 [2]上海市计划生育科学研究所,上海200032

出　　处：《生殖与避孕》2006年第5期259-264,274,共7页Reproduction and Contraception

基　　金：国家自然科学基金资助项目(NO.30472087);上海市科技重大攻关项目(NO.05DZ19314)

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。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： Wes：ern blot results suggested that the expressed proteins with molecular weights of about 125 kD and 138 kD eacn 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.

关 键 词：hCGβ-CTP多聚体 C3D 毕赤酵母 

分 类 号：Q78[生物学—分子生物学]