| 核衣壳蛋白VP39融合TAT转导肽对杆状病毒转导哺乳动物细胞的影响 |
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| 引用本文： | 李俪,王鑫,尹隽,钟江.核衣壳蛋白VP39融合TAT转导肽对杆状病毒转导哺乳动物细胞的影响[J].生物工程学报,2009,25(10):1558-1563. |
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| 作者姓名： | 李俪 王鑫 尹隽 钟江 |
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| 作者单位： | 复旦大学生命科学学院微生物学与微生物工程系,上海,200433 |
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| 基金项目： | 国家自然科学基金项目(No.30770079)资助~~ |
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| 摘 要： | 为了提高昆虫杆状病毒在哺乳动物细胞中转导基因的效率,构建了重组杆状病毒AcRed-tat和AcRed。两者都能在哺乳动物细胞内表达红色荧光蛋白作为报告基因。同时,AcRed-tat带有HIV-1Tat转导肽、病毒主要衣壳蛋白基因vp39及增强型绿色荧光蛋白(egfp)三者的融合基因,并由杆状病毒多角体启动子表达,能够在昆虫细胞中表达该Tat融合蛋白,并掺入子代病毒粒子。而AcRed作为相应的对照病毒,带有多角体启动子表达vp39和egfp的融合基因。2株病毒分别转导哺乳动物细胞后,利用流式细胞仪检测报告基因的表达水平,发现在CHO和Vero细胞中AcRed-Tat介导的报告基因表达水平明显高于AcRed,而在HEK293细胞中2株病毒介导的报告基因表达水平差异不显著。结果表明Tat转导肽可以提高杆状病毒对一部分哺乳动物细胞的转导效率,为改进杆状病毒-哺乳动物细胞转导载体提供了新的思路。
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| 关 键 词： | 杆状病毒 哺乳动物细胞 转导 Tat转导肽 |
| 收稿时间： | 2009/3/10 0:00:00 |
Tat transduction peptide displayed on the nucleocapsid improved the baculovirus transduction of mammalian cells |
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| Li Li,Xin Wang,Juan Yin and Jiang Zhong.Tat transduction peptide displayed on the nucleocapsid improved the baculovirus transduction of mammalian cells[J].Chinese Journal of Biotechnology,2009,25(10):1558-1563. |
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| Authors: | Li Li Xin Wang Juan Yin and Jiang Zhong |
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| Institution: | Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China;Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China;Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China;Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China |
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| Abstract: | In order to improve the transduction efficiency of insect baculovirus in mammalian cells, we constructed two recombinant baculoviruses, AcRed-tat and AcRed. Both viruses expressed red fluorescence protein gene (dsRed) as a reporter in mammalian cell lines. AcRed-tat also contained the coding sequence of HIV-1 Tat transduction peptide fused with viral major capsid protein gene vp39 and enhanced green fluorescence gene (egfp) driven by virus polyhedrin promoter. It expressed the Tat fusion protein in infected... |
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| Keywords: | baculovirus mammalian cells transduction Tat transduction peptide |
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