| DEK真核表达载体的构建及其对p53启动子活性的影响 |
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| 引用本文： | 刘婕,闫志风,张亚楠,林娅红,丁丽华,叶棋浓.DEK真核表达载体的构建及其对p53启动子活性的影响[J].生物技术通讯,2013(1):34-36. |
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| 作者姓名： | 刘婕 闫志风 张亚楠 林娅红 丁丽华 叶棋浓 |
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| 作者单位： | 1. 军事医学科学院 生物工程研究所,北京 100850
2. 解放军总医院 妇产科,北京 100853 |
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| 基金项目： | 国家自然科学基金(31071174) |
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| 摘 要： | 目的：构建DEK的pcDNA3-Flag表达载体，研究其对抑癌基因p53启动子活性的影响。方法：以乳腺文库为模板，PCR扩增DEK编码序列，克隆到pcDNA3-Flag载体，构建成pcDNA3-Flag-DEK，转染293T细胞，Western印迹鉴定peDNA3-Flag载体介导的DEK的表达，萤光素酶报告基因活性实验研究DEK对p53启动子活性的影响。结果：双酶切实验证实得到pcDNA3-Flag-DEK阳性克隆；Western印迹实验发现DEK在293T细胞内表达；转录活性实验表明在ZR75-1乳腺癌细胞中，DEK呈剂量依赖性抑制p53启动子的活性。结论：构建了DEK的真核表达载体，并发现此表达载体能在ZR75-1乳腺癌细胞中抑制p53启动子活性。
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| 关 键 词： | DEK 萤光素酶报告基因 p53启动子 |
Construction of the DEK Eukaryotic Expression Vector and its Effect on p53 Promoter Activity |
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| LIU Jie,YAN Zhi-Feng,ZHANG Ya-Nan,LIN Ya-Hong,DING Li-Hua,YE Qi-Nong.Construction of the DEK Eukaryotic Expression Vector and its Effect on p53 Promoter Activity[J].Letters in Biotechnology,2013(1):34-36. |
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| Authors: | LIU Jie YAN Zhi-Feng ZHANG Ya-Nan LIN Ya-Hong DING Li-Hua YE Qi-Nong |
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| Institution: | 1.Beijing Institute of Biotechnology,Beijing 100850;2.Department of Gynaecology and Obstetrics,Chinese PLA General Hospital,Beijing 100853;China |
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| Abstract: | Objective： To construct a eukaryotic expression vector for expression of DEK, and to study its effect on p53 promoter activity. Methods： The coding sequences of full length DEK was amplified from breast library by PCR and cloned into the pcDNA3-Flag vector. DEK expression was detected by Western blot after the recombi- nant plasmids were transfected into 293T cells. The activity of p53 promoter was detected in ZR75-1 cells. Re- sults： The full length DEK was expressed in the 293T cells. Compared to the empty vector, the activity of p53 promoter was decreased by DEK in a dose-dependent manner. Conclusion： We have constructed the eukaryotic ex- pression vector of DEK and found that p53 promoter activity was decreased by DEK in ZR75-1 cells. |
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| Keywords: | DEK luciferase reporter gene p53 promoter |
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