| Xfect介导重组质粒pEGFP-C2/LAT-ORF3高效转染Vero细胞的方法及其条件的优化 |
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| 引用本文： | 薛礼长,杨慧兰.Xfect介导重组质粒pEGFP-C2/LAT-ORF3高效转染Vero细胞的方法及其条件的优化[J].生物技术通报,2011(1). |
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| 作者姓名： | 薛礼长 杨慧兰 |
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| 作者单位： | 1. 华南理工大学生物科学与工程学院,广州,510006
2. 中国人民解放军广州军区广州总医院,广州,510010 |
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| 基金项目： | 国家自然科学基金资助项目(30972666); 全军医学科学技术研究“十一五”计划资助项目(06J008) |
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| 摘 要： | 旨在用Xfect试剂介导重组质粒pEGFP-C2/LAT-ORF3转染Vero细胞,以期获得转染效率较高的方法,并检测目标片段的表达,从而为进一步研究HSV-2 LAT基因及其ORF3片段的生物学功能奠定基础。用Xfect试剂介导重组质粒pEGFP-C2/LAT-ORF3转染Vero细胞,48 h后观察荧光表达情况,并计算不同转染方法分别在有无血清及质粒与Xfect Polymer比例不同时的转染效率。用RT-PCR检测ORF3片段在Vero细胞中的表达。结果显示,采用贴壁转染法,在有血清及质粒与Xfect Polymer比例为5μg/2μL时转染效率较高;RT-PCR可以获得目的条带,证明ORF3片段在Vero细胞中得到表达。本试验获得的优化条件可以显著提高Xfect Polymer对Vero细胞的转染效率;以绿色荧光蛋白作为标签鉴定目的基因在细胞中表达的方法切实可行。
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| 关 键 词： | Xfect 转染效率 Vero细胞 绿色荧光蛋白 |
Establishment and Optimization of Eukaryotic Expression Plasmid pEGFP-C2/LAT-ORF3 Transfected into Vero Cell by Xfect as Mediation |
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| Xue Lizhang,Yang Huilan.Establishment and Optimization of Eukaryotic Expression Plasmid pEGFP-C2/LAT-ORF3 Transfected into Vero Cell by Xfect as Mediation[J].Biotechnology Bulletin,2011(1). |
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| Authors: | Xue Lizhang Yang Huilan |
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| Institution: | Xue Lizhang1 Yang Huilan2(1School of Biological Science and Engineering,South China University of Technology,Guangzhou 510006,2Guangzhou General Hospital of Guangzhou Military Command,Guangzhou 510010) |
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| Abstract: | In order to obtain highly transfection efficiency and detect the expression of target fragment,the recombinant plasmid pEGFP-C2/LAT-ORF3 was transfected into Vero cells by Xfect as mediation.The recombinant plasmid pEGFP-C2/LAT-ORF3 was transfected into Vero cells mediated by Xfect.After 48 hours observing expression of the recombinant plasmid in the cells by fluorescence microscope,the transfection efficiency was calculated by using transfection methods with yes-no blood serum and different proportions of ... |
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| Keywords: | Xfect Transfection efficiency Vero cells Green fluorescent protein |
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