| 脆性组氨酸三联体基因与HIV-TAT蛋白转导域融合蛋白表达、纯化和多克隆抗体制备 |
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| 引用本文： | 赵亚,周景师,江少杰.脆性组氨酸三联体基因与HIV-TAT蛋白转导域融合蛋白表达、纯化和多克隆抗体制备[J].第四军医大学学报,2009,30(9):831-834. |
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| 作者姓名： | 赵亚 周景师 江少杰 |
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| 作者单位： | 赵亚,ZHAO Ya(第四军医大学,基础部病原生物学教研室,陕西,西安,710033);周景师,ZHOU Jing-Shi(第四军医大学,西京医院肝胆外科,陕西,西安,710033);江少杰,JIANG Shao-Jie(沈阳军区沈阳总医院肝胆外科,辽宁,沈阳,110015)
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| 摘 要： | 目的：制备脆性组氨酸三联体（FHIT）基因与HIV-TAT蛋白转导域的融合蛋白FHIT/TAT以及该融合蛋白的兔抗多克隆抗体,探讨FHIT基因对肝肿瘤的作用及其机制.方法：构建FHIT基因与HIV-TAT蛋白转导域的融合蛋白表达载体,确定该载体的最佳诱导表达条件,在大肠杆菌中大量表达并获得纯化的FHIT/TAT融合蛋白,同时以该融合蛋白免疫兔制备抗FHIT/TAT融合蛋白的多克隆抗体.结果：构建了FHIT基因与HIV-TAT蛋白转导域的融合蛋白表达载体;确定了该载体的最佳诱导表达条件;Western Blot检测也证实了该融合蛋白在大肠杆菌中的表达;以此为基础在大肠杆菌中大量表达并获得了70%纯度的FHIT/TAT融合蛋白;同时制备了较高滴度的兔抗FHIT/TAT融合蛋白多克隆抗体.结论：成功建立了制备具有高效转导作用的HIV-TAT蛋白转导域与FHIT基因融合蛋白的原核表达体系,获得了该融合蛋白的兔抗多克隆抗体,为FHIT基因对于肝肿瘤的作用及其机制研究,特别是肝肿瘤的基因治疗和预防等打下了良好的基础.
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| 关 键 词： | 脆性组氨酸三联体基因 蛋白转导域 基因治疗 |
Studies on expression and purification of FHIT/TAT fusion protein and preparation of its polyclonal antibody |
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| ZHAO Ya,ZHOU Jing-Shi,JIANG Shao-Jie.Studies on expression and purification of FHIT/TAT fusion protein and preparation of its polyclonal antibody[J].Journal of the Fourth Military Medical University,2009,30(9):831-834. |
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| Authors: | ZHAO Ya ZHOU Jing-Shi JIANG Shao-Jie |
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| Institution: | ZHAO Ya1,ZHOU Jing-Shi2,JIANG Shao-Jie31Department of Parasitology,School of Basic Medince,2Department of Hepatobiliary Surgery,Xijing Hospital,Fourth Military Medical University,Xi'an 710033,China,3Department of Hepatobiliary Surgery,PLA Shenyang General Hospital,Shenyang Military Area Command,Shenyang 110015 |
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| Abstract: | AIM:To prepare fusion protein of fragile histidine triad (FHIT)gene with HIV-TAT protein transduction domain and to acquire rabbit polyclonal antibody against the fusion protein so as to lay a good foundation for further research on the function and mechanism of FHIT gene in hepatocellular carcinoma. METHODS: Prokaryotic expression vector of FHIT gene was constructed with HIV-TAT protein transduction domain and its optimal expression conditions were investigated. FHIT/TAT fusion protein was acquired by E.co... |
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| Keywords: | fragile histidine triad(FHIT) gene protein transduction domain(PTD) gene therapy |
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