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| 人Egr-1启动子的克隆及其在肿瘤细胞中的辐射诱导反应 | |
| 引用本文： | 徐小洁,丁丽华,王凌雪,秦玺,程龙,蒋凯,叶棋浓. 人Egr-1启动子的克隆及其在肿瘤细胞中的辐射诱导反应[J]. 细胞与分子免疫学杂志, 2009, 25(11) |
| 作者姓名： | 徐小洁 丁丽华 王凌雪 秦玺 程龙 蒋凯 叶棋浓 |
| 作者单位： | 军事医学科学院,生物工程研究所,北京,100850 |
| 基金项目： | 全军医药卫生科研基金，国家重大科学研究计划 |
| 摘 要： | 目的:克隆人Egr-1基因的启动子, 插入荧光素酶报告基因载体中, 并检测电离辐射对其活性的影响.方法:采用PCR技术从人乳腺癌细胞系MCF-7基因组中扩增出Egr-1启动子, 将其克隆到pGL3-basic载体中;将重组质粒转染人肿瘤细胞, 测定Egr-1启动子在不同辐射条件下转录活性的改变.结果:成功构建了Egr-1启动子的荧光素酶报告基因;在不同剂量的γ射线照射后, Egr-1的启动子活性均明显高于未照射组;在同一剂量照射后48 h, Egr-1的启动子活性达峰值.结论:本实验构建的Egr-1启动子具有辐射激活的功能, 为进一步研究放射-基因治疗奠定了基础. |
| 关 键 词： | Egr-1启动子 辐射 启动子活性 |
Construction of human Egr-1 promoter and its response to ionizing radiation in tumor cells |
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| XU Xiao-jie,DING Li-hua,WANG Ling-xue,QIN Xi,CHENG Long,JIANG Kai,YE Qi-nong. Construction of human Egr-1 promoter and its response to ionizing radiation in tumor cells[J]. Chinese journal of cellular and molecular immunology, 2009, 25(11) | |
| Authors: | XU Xiao-jie DING Li-hua WANG Ling-xue QIN Xi CHENG Long JIANG Kai YE Qi-nong |
| Abstract: | AIM: To construct human Egr-1 promoter luciferase reporter system and study its activity induced by i-onizing radiation. METHODS: Egr-1 promoter was obtained by human genomic PCR and cloned into pGL3-basic vector. After transfection of recombinant plasmid into human tumor cells, the Egr-1 promoter activity induced by ionizing radiation was detected by luciferase reporter assay. RESULTS: The luciferasy reporter system of Egr-1 promoter was successfully constructed. The activity of Egr-1 promoter was substantially increased after different doses of IR and reached to the peak at the time point of 48h after IR. CONCLUSION: The Egr-1 promoter was constructed in this study showed IR inducible activity in tumor cells, laying foundation for the research of radiation, mediated gene therapy. |
| Keywords: | Egr-1 promoter ionizing radiation promoter activity |
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