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| 影响生长抑素重组质粒pEGS/2SS转染HeLa细胞效果的因素研究 | |
| 引用本文： | 何晓红,曹少先,茆达干,杨利国.影响生长抑素重组质粒pEGS/2SS转染HeLa细胞效果的因素研究[J].畜牧兽医学报,2010,41(3). |
| 作者姓名： | 何晓红 曹少先 茆达干 杨利国 |
| 作者单位： | 1. 南京农业大学动物科技学院,南京,210095;中国农业科学院北京畜牧兽医研究所,北京,100193 2. 江苏省农业科学院畜牧研究所,南京,210014 3. 南京农业大学动物科技学院,南京,210095 4. 华中农业大学动物科技学院,武汉,430070 |
| 摘 要： | 旨在分析生长抑素重组质粒pEGS/2SS转染HeLa细胞(人宫颈癌上皮细胞)转染效果的影响因素,探讨最佳转染条件,为进一步研究生长抑素基因疫苗的作用机制和作用效果奠定基础。以脂质体转染法将带有绿色荧光蛋白(GFP)报告基因的重组质粒pEGS/2SS转染HeLa细胞,探讨质粒转染HeLa细胞最佳条件的4个参数:质粒DNA剂量、脂质体剂量、最佳转染时间和质粒提取方法,在荧光显微镜下观察细胞转染情况并计算转染率。结果:在六孔细胞培养板上,当DNA/脂质体剂量为1μg/6μg时,转染效率最高,在转染后72 h达到34.02%;当用Plasmid Maxi Kit试剂盒提取质粒,转染时间为48 h,转染效率最高,达到17.88%。重组质粒pEGS/2SS转染He-La细胞条件是:采用试剂盒(Plasmid Maxi Kit)提取的质粒,DNA和脂质体的剂量分别为1μg与6μg,转染时间48 h,此时的转染效率最高,达17.88%。 |
| 关 键 词： | 脂质体 HeLa细胞 生长抑素重组质粒pEGS/2SS |
Transient Transfection Factors for Expression of Recombinant Somatostatin Plasmid pEGS/2SS in Cultured HeLa Cells |
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| HE Xiao-hong,CAO Shao-xian,MAO Da-gan,YANG Li-guo.Transient Transfection Factors for Expression of Recombinant Somatostatin Plasmid pEGS/2SS in Cultured HeLa Cells[J].Acta Veterinaria et Zootechnica Sinica,2010,41(3). | |
| Authors: | HE Xiao-hong CAO Shao-xian MAO Da-gan YANG Li-guo |
| Institution: | HE Xiao-hong1,4,CAO Shao-xian2,MAO Da-gan1,YANG Li-guo3(1.College of Animal Science , Technology,Nanjing Agricultural University,NanJing210095,China,2.Institute of Animal Science,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,3.College of Animal Science , Technology,HuazhongAgricultural University,Wuhan 430070,4.Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing 100193,China) |
| Abstract: | We aimed to investigate the factors infecting transfection efficiency of somatostatin recombinant plasmid pEGS/2SS introducting into HeLa cells,make preparations for advanced study on somatostatin gene vaccine mechanism and effect of action.Four parameters including transfection reagent(lipofectamineTM) concentration,DNA concentration,cells cultured time after transfection and DNA extracting techniques were analyzed and optimized,and transfection efficiency was evaluated by the percentage of transfected cel... |
| Keywords: | liposome HeLa cells somatostatin recombinant plasmid pEGS/2SS |
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