| 人BclGL基因慢病毒表达载体构建及其对T淋巴细胞凋亡的影响 |
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| 引用本文： | 罗娜,吴毅,费蕾,郭晟,杨承英,吴胜昔,陈永文,吴玉章,郝飞. 人BclGL基因慢病毒表达载体构建及其对T淋巴细胞凋亡的影响[J]. 免疫学杂志, 2009, 25(3) |
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| 作者姓名： | 罗娜 吴毅 费蕾 郭晟 杨承英 吴胜昔 陈永文 吴玉章 郝飞 |
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| 作者单位： | 罗娜,郝飞(第三军医大学,西南医院皮肤科);吴毅(第三军医大学,基础部解剖教研室);费蕾,郭晟,杨承英,吴胜昔,陈永文,吴玉章(第三军医大学,基础部全军免疫学研究所) |
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| 摘 要： | 目的 构建针对人BclGL的慢病毒表达载体,检测其对Jurkat T淋巴细胞系凋亡的影响,为进一步研究BclGL在细胞内信号转导及分子功能奠定基础.方法 RT-PCR方法扩增人全长BclGL基因,克隆于pGrP-N1质粒中构建BclGL绿色荧光蛋白融合基因(BclGL-GFP),将融合基因克隆于慢病毒载体FUGW并转染293FT细胞包装浓缩慢病毒颗粒.将浓缩慢病毒感染Jurkat细胞,同时设立PBS对照及FUGW病毒阴性对照组,流式细胞术及荧光定量PCR鉴定BclGL在Jurkat细胞中的表达,Annexin V-APC/PI双染检测细胞凋亡变化.结果 酶切及测序证实人BclGL慢病毒表达载体构建成功,转染Jurkat细胞72 h后Jurkat细胞凋亡明显增加.结论成功构建了高效稳定表达人BclGL的重组慢病毒转染系统,并证实其对Jurkat淋巴细胞的促凋亡效应,为进一步探讨BclGL的生物学功能奠定了基础.
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| 关 键 词： | 慢病毒表达载体 Jurkat细胞 凋亡 |
Recombinant lentivirus vector construction of human BclGL and its influence on Jurkat cell apoptosis |
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| LUO Na,WU Yi,FEI Lei,GUO Sheng,YANG Chengying,WU Shengxi,WU Yuzhang,HAO Fei. Recombinant lentivirus vector construction of human BclGL and its influence on Jurkat cell apoptosis[J]. Immunological Journal, 2009, 25(3) |
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| Authors: | LUO Na WU Yi FEI Lei GUO Sheng YANG Chengying WU Shengxi WU Yuzhang HAO Fei |
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| Affiliation: | LUO Na,WU Yi,FEI Lei,GUO Sheng,YANG Chengying,WU Shengxi,WU Yuzhang,HAO Fei Department of Dermatology,Southwest Hospital,Third Military Medical University,Chongqing,400038,China |
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| Abstract: | Objective To construct a recombinant lentivirus vector encoding proapoptotic protein BclGL and investigate its effects on Jurkat cell apoptosis,for further studng of apoptotic signal transduction. Methods Full-length human-derived BclGL DNA,obtained by PCR,was inserted into pGFP-N1 vector to construct fusion gene BclGL-GFP,which then was amplified by PCR and inserted into FUGW vector after endonuclease digestion. The fusion gene was identified by restriction endonuclease digestion and nucleotide sequencing.... |
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| Keywords: | BclGL |
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