| 人精子蛋白17增强型绿色荧光蛋白共表达卵巢癌细胞模型的建立 |
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| 引用本文： | 韩艳玲,李芳秋,刘群,周万青,张蕾,王立魁.人精子蛋白17增强型绿色荧光蛋白共表达卵巢癌细胞模型的建立[J].医学研究生学报,2008,21(9). |
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| 作者姓名： | 韩艳玲 李芳秋 刘群 周万青 张蕾 王立魁 |
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| 作者单位： | 南京军区南京总医院解放军临床检验医学研究所,江苏南京,210002 |
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| 基金项目： | 国家自然科学基金，南京军区医学科技研究计划重点项目，国家重点基础研究发展计划(973计划) |
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| 摘 要： | 目的:建立人精子蛋白17(hSp17)与增强型绿色荧光蛋白(EGFP)共表达的卵巢癌细胞模型. 方法:用PCR方法获取hSp17基因片段,Hind Ⅲ/Kpn I双酶切、T4 DNA连接酶连接的方法将hSp17基因片段克隆至含报告基因EGFP的真核表达载体pEGFP-N1中,脂质体介导转染卵巢癌细胞HO8910,荧光倒置显微镜观察及Westernblot检测转染细胞中hSp17/EGFP融合蛋白的表达,G418筛选稳定表达的细胞株. 结果:酶切和测序结果均证实pEGFP-N1/hSp17重组质粒的构建完全正确.荧光观察与Western blot均证实了hSp17/EGFP蛋白的共表达,且成功筛选出稳定表达的细胞株. 结论:成功构建hSp17/EGFP共表达的卵巢癌细胞模型,为研究hSp17异常表达对肿瘤细胞生物学行为的影响,以及hSp17为靶标的肿瘤诊断和生物治疗奠定基础.
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| 关 键 词： | 人精子蛋白17 卵巢癌 绿色荧光蛋白 |
Establishment of an ovarian carcinoma cell model with co-expression of human sperm protein 17 and enhanced green fluorescent protein |
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| HAN Yan-ling,LI Fang-qiu,LIU Qun,ZHOU Wan-qing,ZHANG Lei,WANG Li-kui.Establishment of an ovarian carcinoma cell model with co-expression of human sperm protein 17 and enhanced green fluorescent protein[J].Bulletin of Medical Postgraduate,2008,21(9). |
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| Authors: | HAN Yan-ling LI Fang-qiu LIU Qun ZHOU Wan-qing ZHANG Lei WANG Li-kui |
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| Institution: | HAN Yan-ling,LI Fang-qiu,LIU Qun,ZHOU Wan-qing,ZHANG Lei,WANG Li-kui(PLA Institute of Clinical Laboratory Medicine,Nanjing General Hospital of Nanjing Military Comm,,PLA,Nanjing 210002,Jiangsu,China) |
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| Abstract: | Objective: To establish a human ovarian carcinoma(OC) cell model that stably co-expresses the human sperm protein 17 and enhanced green fluorescent protein(hSp17/EGFP).Methods: hSp17 cDNA was obtained by PCR and cloned into the vector pEGFP-N1 that contained a report gene-enhanced green fluorescent protein.The recombinant plasmid was transfected into HO8910 cells with lipofectamine 2000.The expression of fusion hSp17/EGFP was observed under the fluorescent inverted microscope and detected by Western blottin... |
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| Keywords: | Sperm protein 17 Ovarian cancer Green fluorescent protein |
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