| hCGb-CTP多聚体-3C3d融合蛋白在巴氏毕赤酵母中的表达 |
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| 引用本文： | 冯文华,石燕,喻光荣,沈卫英,蒋俶,申庆祥.hCGb-CTP多聚体-3C3d融合蛋白在巴氏毕赤酵母中的表达[J].生殖与避孕,2006(5). |
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| 作者姓名： | 冯文华 石燕 喻光荣 沈卫英 蒋俶 申庆祥 |
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| 作者单位： | 复旦大学上海医学院，复旦大学上海医学院，复旦大学上海医学院，上海市计划生育科学研究所，上海市计划生育科学研究所，上海市计划生育科学研究所 上海，200032上海市计划生育科学研究所，上海，200032，上海，200032上海市计划生育科学研究所，上海，200032，上海，200032上海市计划生育科学研究所，上海，200032，上海，200032，上海，200032，上海，200032 |
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| 基金项目： | 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) |
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| 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。
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| 关 键 词： | hCGβ-CTP多聚体 C3d 毕赤酵母 |
Expression of Fusion Protein hCGβ-CTP Polymer and 3C3d in Pichia pastoris Cells |
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| Wen-hua FENG,Yan SHI,Guang-rong YU,Wei-ying SHEN,Shu JIANG,Qing-xiang SHEN.Expression of Fusion Protein hCGβ-CTP Polymer and 3C3d in Pichia pastoris Cells[J].Reproduction and Contraception,2006(5). |
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| Authors: | Wen-hua FENG Yan SHI Guang-rong YU Wei-ying SHEN Shu JIANG Qing-xiang SHEN |
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| Institution: | Wen-hua FENG1,2,Yan SHI1,2,Guang-rong YU1,2,Wei-ying SHEN2,Shu JIANG2,Qing-xiang SHEN2 |
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| 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. |
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| Keywords: | hCGβ-CTP polymer C3d Pichia pastoris yeast |
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