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低氧诱导人VEGF基因表达载体的构建及其蛋白表达检测 被引量：2

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摘要 目的以人血管内皮细胞生长因子(hVEGF)基因的低氧反应元件(HRE)作为增强子,构建含HRE的真核表达载体,探讨含HRE不同拷贝数的载体对低氧的反应状态。方法将人工合成的HRE串联后与CMV启动子连接,替换pcDNA3.1的启动子部分,构建成低氧诱导真核表达载体;将hVEGF基因重组到此载体中,转染BHK细胞。用免疫组化染色法检测VEGF的表达,用酶联免疫吸附实验(ELISA)检测低氧前后细胞hVEGF表达的变化。结果成功构建了含HRE的低氧诱导真核表达载体4HRE-pcDNA-VEGF和6HRE-pcDNA-VEGF。转染细胞后进行低氧培养,VEGF的表达均增高,含6HRE的低氧诱导作用高于4HRE。结论在含HRE的真核表达载体中,低氧时可诱导基因的表达,其拷贝数可影响诱导作用的强度。
作者 殷爱红 武文琦 何丽明 郑少鹏
机构地区 首都医科大学
出处 《山东医药》 CAS 北大核心 2011年第52期48-50,I0002,共4页 Shandong Medical Journal
基金 北京市教育委员会资助项目(KM200310025094)
关键词 缺氧诱导因子 低氧反应元件 真核表达载体 血管内皮细胞生长因子
分类号 R34 [医药卫生—基础医学] Q782 [生物学—分子生物学]
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参考文献6

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共引文献4

同被引文献27

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引证文献2

二级引证文献1

山东医药
2011年 第52期

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