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丙型肝炎病毒核心蛋白酵母双杂交诱饵载体的构建及人胎肝cDNA文库的扩增、纯化和鉴定
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【医学免费毕业论文】作者：刘敏，张伟，陈天艳，蔺淑梅，赵良，刘锦锋，赵英仁，张树林【关键词】 丙型肝炎病毒核心蛋白摘要：目的 用含丙型肝炎病毒核心(HCV core)蛋白的cDNA片段构建酵母双杂交诱饵载体，并进行人胎肝cDNA文库的扩增、纯化和鉴定。 方法 PCR扩增含HCV core蛋白不同大小的cDNA片段，分别克隆入pUC19质粒，经测序正确后，再亚克隆入酵母双杂交诱饵载体pGBKT7中。扩增人胎肝cDNA文库并纯化、鉴定。结果 获得含HCV core蛋白的cDNA片段，并成功克隆入pGBKT7中。待转化的人肝cDNA文库滴度在5×108左右，纯化后的质粒 DNA质量浓度约1g/L。用EcoRⅠ、XhoⅠ双酶切显示插入片段大小不一。结论 成功构建了核心蛋白的酵母双杂交诱饵载体；扩增、纯化的人胎肝cDNA文库的多样性很好，适合于筛选。为用酵母双杂交技术 研究 与HCV core蛋白相互作用的蛋白打下坚实基础。
关键词：丙型肝炎病毒核心蛋白；酵母双杂交；cDNA文库
ABSTRACT: Objective To construct the bait vector of HCV core region in yeast twohybrid system, and to amplify, purify and evaluate the cDNA gene bank of human fetal liver. Methods The cDNA fragments encoding HCV core region were amplified by PCR, and then were cloned into pUC19. After being verified by sequencing, they were subcloned into the bait vector pGBKT7 of yeast twohybrid system, subsequently amplified and purified. And then the cD
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