| hCGβ-C3d融合蛋白在CHO细胞中的表达 |
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| 引用本文： | 王明雁,李磊,李大金,申庆祥,朱影.hCGβ-C3d融合蛋白在CHO细胞中的表达[J].现代免疫学,2003,23(1):13-15,39. |
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| 作者姓名： | 王明雁 李磊 李大金 申庆祥 朱影 |
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| 作者单位： | 1. 复旦大学附属妇产科研究所生殖免疫研究室,上海,200011
2. 上海市计划生育研究所,上海,200032 |
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| 基金项目： | 国家自然科学基金资助项目 (No .3 0 2 712 3 5 )，计划生育药具国家重点实验室资助项目 (No B2 0 1 1)，上海市自然科学基金资助项目 (No 0 0ZB14 0 5 8)，教育部博士学科点专项科研基金资助项目 (No 9941)，卫生部优秀青年人才专项科研基金资助项目(No 97 |
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| 摘 要： | 为了提高hCGβ的免疫原性 ,构建含新型分子佐剂C3d的hCGβ真核细胞表达质粒pcDNA3 hCGβ C3d3,使其转染稳定表达系统中国仓鼠卵细胞 (CHO )后 ,检测融合蛋白分泌情况。CHO细胞转染了pcDNA3 hCGβ C3d3质粒后 ,经 4 8h培养 ,1 0× 10 6个CHO细胞可分泌 6 6 0ng的重组融合hCGβ C3d3蛋白。细胞免疫化学技术分析显示 ,表达产物既有hCGβ的组分也有C3d的成分。经SDS PAGE及免疫印迹试验发现CHO细胞培养上清液中含有三组蛋白成分 ,其相对分子质量分别为12 0 0 0 0、90 0 0 0和 6 5 0 0 0。利用新型分子佐剂C3d与hCGβ的连接 ,构建了pcDNA3 hCGβ C3d3真核细胞表达质粒 ,为提高hCGβDNA免疫避孕疫苗的免疫原性及抗生育作用奠定了基础
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| 关 键 词： | hCGβ C3d3 CHO Raji细胞 |
| 文章编号： | 1001-2478(2003)01-0013-03 |
Expression of hCGβ-C3d Fusion Protein in CHO Cells |
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| WANG Ming yan ,LI Lei ,LI Da jin ,SHEN Qing xiang ,ZHU Ying.Expression of hCGβ-C3d Fusion Protein in CHO Cells[J].Current Immunology,2003,23(1):13-15,39. |
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| Authors: | WANG Ming yan LI Lei LI Da jin SHEN Qing xiang ZHU Ying |
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| Institution: | WANG Ming yan 1,LI Lei 1,LI Da jin 1,SHEN Qing xiang 2,ZHU Ying 1 |
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| Abstract: | To construct a new plasmid to enhance the immunogenicity of hCGβ with potent molecular adjuvant C3d,a plasmid pcDNA3 hCGβ C3d3 was constructed with the fusion protein determined and analyzed from transfected CHO cells by methods of immunohistochemistry and Western blotting It was found that the stable expression system of CHO cells transfected by this plasmid could secret the recombinant fusion protein hCGβ C3d3 at a speed of 660 ng/10 6 cells/48 hrs The expression products contained both C3d receptor binding activity and hCGβ antigenicity as detected by Raji cell immunohistochemical assay There appeared as 3 protein bands in the supernatants of cultures of transfected cells bearing anti hCGβ activity on SDS PAGE and Western blots with molecular weight of 120,90 and 65 kD respectively It concludes that we have successfully constructed a plasmid pcDNA3 hCGβ C3d3 that is able to express the fusion proteins in eukaryotic cells and may provide a useful tool for further researches in hCGβ contraceptive DNA and protein vaccines |
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| Keywords: | C3d3 CHO |
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