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淀粉样前体蛋白的原核表达、纯化及抗血清的制备和应用 被引量：1

Expression and Purification of Recombinant Human APP Proteins in E.coli and Preparation of Rabbit Antiserum Against APP
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摘要 提取SH-SY5Y细胞的总RNA并合成单链cDNA,用PCR方法扩增出APP基因羧基端片段.测序后克隆至表达载体,在大肠杆菌M15中分别表达APP-C100和GST-APP-C100融合蛋白,纯化的GST-APP-C100分子量约为38kD,APP-C100约为18kD,此外在APP-C100纯化产物中还存在有超过97kD的聚合物.体外蛋白酶消化实验显示纯化的GST-APP-C100和APP-C100单体对蛋白酶K消化敏感,而APP-C100聚合物具有抵抗蛋白酶消化能力.以GST-APP-C100免疫实验用兔后得到的APP抗血清经Western印迹鉴定可特异性识别重组及细胞内源性APP.
作者 石琦 韩俊 高晨 李锋 张宝云 周玉玲 董小平
机构地区 中国疾病预防控制中心病毒病预防控制所传染病预防控制国家重点实验室 西安交通大学医学院
出处 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2006年第3期258-261,共4页 Chinese Journal of Biochemistry and Molecular Biology
基金 国家自然科学基金委重点项目(No.30130070) 国家高技术研究发展计划(863计划)项目(No.2001AA215391) 欧盟项目(QLRT200001441)和国家科技攻关计划项目(No.2003BA712A04-02)资助~~
关键词 淀粉样前体蛋白 原核表达 蛋白酶抗性 多克隆抗体
分类号 R392.1 [医药卫生—免疫学]
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参考文献12

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共引文献5

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中国生物化学与分子生物学报
2006年 第3期

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