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狂犬病病毒HEP-Flury株N基因的修饰及原核表达

DOI: 10.7671/j.issn.1001-411X.2012.01.021, PP. 102-105

Keywords: 狂犬病病毒,核蛋白,原核表达

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Abstract:

为了高效表达狂犬病病毒HEP-Flury的核蛋白,选取抗原决定簇富集区,利用生物信息学技术分析了狂犬病病毒HEP-FluryN蛋白的核苷酸序列。根据大肠埃希菌Escherichiacoli对密码子的偏嗜性,首先对所选取的核苷酸序列进行修饰、引入酶切位点,然后定向克隆至原核表达载体pET-32a(+),构建重组质粒pET-32-NP。将其转化表达宿主菌EscherichiacoliBL21(DE3)pLysS,以IPTG诱导表达.表达产物经SDS-PAGE电泳,发现在相对分子质量34000处有1条特异的蛋白带,与预期大小一致.Western-blotting检测结果显示,该蛋白可被鼠抗His单克隆抗体及犬抗RV多克隆抗体特异识别,表明所表达的N蛋白具有良好的免疫原性。

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