| 通过共表达转铁蛋白和细胞周期蛋白D1改造CHO细胞株 |
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| 引用本文： | 付玲,赵兴卉,郭俊伟,于婷,宋小红,侯利华,陈薇. 通过共表达转铁蛋白和细胞周期蛋白D1改造CHO细胞株[J]. 生物技术通讯, 2012, 0(5): 676-679 |
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| 作者姓名： | 付玲 赵兴卉 郭俊伟 于婷 宋小红 侯利华 陈薇 |
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| 作者单位： | 军事医学科学院 生物工程研究所,病原微生物生物安全国家重点实验室2004DAV00214,北京 100071 |
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| 基金项目： | 国家自然科学基金(81025018,30570951) |
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| 摘 要： | 目的:对现有的CHO-DG44细胞株进行改造,得到在无血清培养基中更具生长优势的CHO宿主细胞株。方法:分别克隆转铁蛋白和细胞周期蛋白D1基因,构建共表达2种基因的pIRES质粒,转染CHO-DG44细胞株,筛选G418抗性克隆。结果与结论:得到3株G418阳性克隆株,其中转铁蛋白和细胞周期蛋白D1表达水平最高的S6与CHO-DG44相比,在无血清培养基中生长更快、密度更高。
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| 关 键 词： | 中国仓鼠卵巢细胞 代谢工程 转铁蛋白 细胞周期蛋白D1 |
Construction of Optimized CHO Cell Lines by Overexpression of Transferrin and Cyclin D1 |
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| FU Ling,ZHAO Xing-Hui,GUO Jun-Wei,YU Ting,SONG Xiao-Hong,HOU Li-Hua,CHEN Wei. Construction of Optimized CHO Cell Lines by Overexpression of Transferrin and Cyclin D1[J]. Letters in Biotechnology, 2012, 0(5): 676-679 |
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| Authors: | FU Ling ZHAO Xing-Hui GUO Jun-Wei YU Ting SONG Xiao-Hong HOU Li-Hua CHEN Wei |
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| Affiliation: | State Key Laboratory of Pathogens and Biosecurity(2004DAV00214),Beijing Institute of Biotechnology,Beijing 100071,China |
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| Abstract: | Objective: To optimize CHO-DG44 cell line for better performance in serum-free culture.Methods: Transferrin and cyclin D1 ORFs were inserted into pIRES,then the construct was transfected into CHO-DG44 cells.The cells were treated with G418 at a final concentration of 800 μg/mL for selection.Results and Coclu sion: Three CHO-DG44 cell lines overexpressing transferrin and cyclin D1 were obtained.The growth rate of one of these three cell lines,S6,was significantly greater than CHO-DG44 during days 4~7 of suspension culture in serum-free medium.Our study proved that overexpression of cyclin D1 and transferrin could confer CHO-DG44 with increased growth rate,and provided new candidate CHO cell lines for biopharmaceutical production. |
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| Keywords: | Chinese hamster ovary cell metabolic engineering transferrin cyclin D1 |
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