| hTRT基因荧光表达载体在真核细胞中的表达 |
| 投稿时间：2010-03-09 修订日期：2010-03-16 点此下载全文 |
| 引用本文：周海文,杨雯君,刘德莉,刘伟,周曾同.hTRT基因荧光表达载体在真核细胞中的表达[J].医学研究杂志,2010,39(5):21-23 |
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| 基金项目:国家自然科学基金（39970791）；上海市教委资助项目（04BC45） |
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| 中文摘要:目的研究具绿色荧光蛋白报道系统的端粒酶基因真核质粒载体在真核细胞中的表达情况。方法用LipofectAMINE 2000介导重组端粒酶pEGFP-hTRT质粒转染293细胞及人正常口腔黏膜细胞，经荧光显微镜、激光共聚焦显微镜观察荧光表达并进行流式细胞仪检测转染效率，采用RT-PCR检测转染细胞中外源性hTRT的表达。结果质粒在293细胞及人正常口腔黏膜细胞中正确表达，外源性端粒酶在细胞中表达。293细胞及人正常口腔黏膜角化细胞转染效率分别为34.74%和6.88%。结论hTRT基因荧光表达载体在真核细胞中表达，便于观察转染情况，对研究组织工程种子细胞的衰老问题具有重要的意义。 |
| 中文关键词:端粒酶 绿色荧光蛋白 基因转染 质粒 |
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| Expression of Recombinant hTRT Plasmid in Eukaryotic Cells |
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| Abstract:ObjectiveTo study the expression of human Telomerase Reverse Transcriptase (hTRT) in-vitro cell cultivation by transfecting plasmid pEGFP-hTRT into eukaryote for exploring. MethodsThe pEGFP-hTRT was transfected into 293 cells and normal human oral keratinocytes(OKC) by lipofectAMINE 2000. Transfection efficiency of the Green Fluorescent Protein (GFP) was detected under fluorescent microscope. Transfection efficiency was examined by flow cytometer. Exogenous hTRT was analyzed by RT-PCR. ResultsTransfeced cells showed green fluorescent. Exogenous hTRT was reconstructed. The transfection efficiency of 293 cells and OKC was 34.74%和6.88% respectively. ConclusionThe pEGFP-hTRT has been successfully transfected into eukaryotic cells, which has an important role for extension life-span of normal human cells. |
| keywords:hTRT Transgenic plasmid GFP Liposome |
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