| HPC2真核表达载体的构建及其在HEK293细胞中的表达 |
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| 引用本文： | 谭康联 李志杰 刘靖华 黄浩 唐靖 邓鹏 姜勇. HPC2真核表达载体的构建及其在HEK293细胞中的表达[J]. 第一军医大学学报, 2005, 25(12): 1482-1484,1497 |
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| 作者姓名： | 谭康联 李志杰 刘靖华 黄浩 唐靖 邓鹏 姜勇 |
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| 作者单位： | [1]南方医科大学广东省功能蛋白质组学重点实验室,广东广州510515 [2]广州中医药大学第二临床医学院,广东广州510405 [3]现为广州中医药大学在读七年制硕士研究生,广东广州510405 |
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| 基金项目： | 国家重点基础研究发展项目（973）（2002CB513000）：国家高技术研究发展计划（863）（2001AA234061）. |
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| 摘 要： | 目的 构建在哺乳动物细胞中表达的HPC2真核表达载体，并分析其在HEK293细胞中的表达。方法 从重组pcDNA3／HPC2载体上将HPC2cDNA序列亚克隆至带有flag标记的真核表达载体pcDNA3-flag上，经PCR、酶切和测序鉴定，将重组质粒pcDNA3-flag／HPC2用脂质体瞬时转染HEK293细胞，细胞裂解后，用Western blot分析并观察HPC2在HEK293细胞中的表达情况。结果 PCR、酶切和DNA测序结果均表明重组质粒pcDNA3-flag／HPC2构建正确，并可在HEK293细胞内高效表达。结论 成功构建了pcDNA3-flag／HPC2真核表达载体，该载体能在哺乳动物细胞中有效表达，为进一步研究HPC2的功能提供了重要的实验材料。
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| 关 键 词： | HPC2 载体构建 基因表达 转染 |
| 文章编号： | 1000-2588（2005）12-1482-03 |
| 收稿时间： | 2005-07-03 |
| 修稿时间： | 2005-07-03 |
Construction of eukaryotic expression vector for HPC2 and its expression in HEK293 cells |
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| TAN Kang-lian , LI Zhi-jie, LIU Jing-hua, HUANG Hao, TANG Jing, DENG Peng, JIANG Yong. Construction of eukaryotic expression vector for HPC2 and its expression in HEK293 cells[J]. Journal of First Military Medical University, 2005, 25(12): 1482-1484,1497 |
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| Authors: | TAN Kang-lian LI Zhi-jie LIU Jing-hua HUANG Hao TANG Jing DENG Peng JIANG Yong |
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| Affiliation: | 1.Key Laboratory of Functional Proteomics of Guangdong Province, Southern Medical University, Guangzhou 510515, China; 2 Second Clinical Medical College, Guangzhou University of Traditional Chinese Medicine, Guangzhou 510405, China |
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| Abstract: | Objective To construct the eukaryotic expression vector for HPC2 for expression in HEK293 cells. Methods HPC2 from pcDNA3/HPC2 were inserted into the flag-tagged vector pcDNA3-flag by subcloning method. The recombinant plasmid pcDNA3-flag/HPC2 was then transfected into HEK293 cells using a routine lipofectamine method. The cell lysate was used for Western blotting to examine the expression of the target protein. Results and Conclusion Double restriction enzyme digestion and DNA sequencing indicated successful construction of the eukaryotic expression vector for HPC2 and the fusion protein was highly expressed in HEK293 cells, which provides an important basis for functional study of HPC2. |
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| Keywords: | HPC2 vector construction gene expression transfection |
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