| 过表达热休克蛋白70提高CHO细胞表达抗体的能力 |
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| 引用本文： | 黄芬,安小平,李存,何彰华,张宝中,米志强,王娟,童贻刚.过表达热休克蛋白70提高CHO细胞表达抗体的能力[J].中国生物工程杂志,2011,31(9):8-13. |
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| 作者姓名： | 黄芬 安小平 李存 何彰华 张宝中 米志强 王娟 童贻刚 |
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| 作者单位： | 1. 军事医学科学院微生物流行病研究所 病原微生物生物安全国家重点实验室 北京 100071;
2. 陕西师范大学生命科学学院 西安 710062 |
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| 基金项目： | 国家自然科学基金，国家"863"计划，病原微生物生物安全国家重点实验室开放课题 |
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| 摘 要： | 通过在中国仓鼠卵巢细胞(CHO)中过表达热休克蛋白70以提高其表达抗体的能力。首先从中国仓鼠基因组DNA中扩取HSP70基因,构建真核表达质粒pcDNA3.1-HSP70,再将重组质粒稳定转染到CHO/dhfr-细胞中,筛选获得稳定的细胞系,运用RT-qPCR检测和Western blot分析HSP70基因的过表达。在过表达HSP70的CHO细胞组和对照细胞组(转染空载体pcDNA3.1的CHO细胞组)中分别转染表达抗-HBs的质粒,应用ELISA检测两组细胞表达抗-HBs的能力。RT-qPCR结果显示实验组CHO细胞中HSP70基因的表达量明显高于对照组细胞;ELISA检测结果表明过表达HSP70的CHO细胞组抗-HBs表达量高于对照组细胞(P<0.05)。研究揭示HSP70能有效促进细胞内分泌性蛋白的表达。
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| 关 键 词： | CHO细胞 HSP70 抗体 |
| 收稿时间： | 2011-05-30 |
| 修稿时间： | 2011-07-01 |
Overexpression of Heat Shock Protein70 Enhances Production of Antibody in Chinese Hamster Ovary Cells |
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| HUANG Fen,AN Xiao-ping,LI Chun,HE Zhang-hua,ZHANG Bao-zhong,MI Zhi-qiang,WANG Juan,TONG Yi-gang.Overexpression of Heat Shock Protein70 Enhances Production of Antibody in Chinese Hamster Ovary Cells[J].China Biotechnology,2011,31(9):8-13. |
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| Authors: | HUANG Fen AN Xiao-ping LI Chun HE Zhang-hua ZHANG Bao-zhong MI Zhi-qiang WANG Juan TONG Yi-gang |
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| Institution: | 1(1 Institute of Microbiology and Epidemiology,Academy of Military Medical Sciences,Beijing 100071,China)(2 College of Life Science,Shaanxi Normal University,Xi’an 710062,China) |
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| Abstract: | The production of recombinant antibody was significantly enhanced by the method of overexpression of Heat Shock Protein 70 in Chinese Hamster Ovary (CHO) cells. The gene HSP70 was cloned from CHO/dhfr-cells, then HSP70 expression plasmid pcDNA3.1-HSP70 was constructed and transfected into CHO/dhfr-cells. CHO cell line overexpressing HSP70 stably was isolated and the expression of HSP70 gene was analyzed through RT-PCR. Anti-HBs chimeric antibody expression plasmids were transfected into the HSP70 cell line, ELISA was performed to test the production of anti-HBs antibody. It demonstrated that the overexpression of HSP70 can enhance the production of recombinant antibody in CHO/dhfr-cells (P<0.05). |
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| Keywords: | Chinese hamster ovary cells Heat Shock Protein 70 Gene expression Antibody |
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