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猪流感病毒抗原表位基因表达载体构建与免疫原性分析 被引量：5

Construction and Immunogenicity Analysis of Antigenic Epitopes of Swine Influenza Virus
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摘要 根据猪流感病毒血凝素蛋白基因(Heamuglutinine,HA)的核苷酸序列,设计、筛选HA蛋白氨基酸序列的主要表位多肽4个,将4个片段以柔性连接串联成模拟蛋白,核苷酸约为300 bp,体外扩增该模拟蛋白基因,插入到原核表达载体pET30a(+)中,转染宿主菌诱导表达,结果获得分子量为20 kD的表达蛋白,该蛋白可与抗His-tag抗体、抗猪流感病毒H1N1、H3N2亚型高免血清发生免疫学反应。纯化后免疫小鼠,ELISA及血凝抑制(Heamuglutinine inhibitor,HI)试验检测,小鼠产生针对多肽抗原的血清抗体,同时还可检测到H1N1、H3N2亚型SIV血凝抗体。流氏细胞仪检测免疫组外周血淋巴细胞高于对照组,说明该模拟蛋白具有与H1N1、H3N2亚型猪流感病毒相似的免疫原性及反应原性,为H1N1、H3N2血清亚型猪流感病毒疫苗研制提供了新手段。 Several antigen epitopes were designed according to the sequences of Swine influenza virus hemagglutinin （HA） genes and lined with the interval. The gene was amplified by PCR and sub cloned into pET30a （＋） vector. The fusion protein was expressed in E. coli BL21 （DE3） by induced with IPTG and purified by affinity chromatography. The molecular weight of the protein was about 20 kD in SDS-PAGE. Immunological activity of the fusion protein was analyzed by Western blot. The results showed that the fusion protein could interact with anti-His antibody and the rabbit antiserum against SIV. The immunized mouse can produced antibodies against the target peptide and HI antibody against SIV H1N1 or H3N2. This study provides a new vaccine candidate for SIV.
作者 刘惠莉 邢继兰 潘洁 杨秋峰 赵艳敏
机构地区 上海市农业科学院畜牧兽医研究所
出处 《生物工程学报》 CAS CSCD 北大核心 2008年第4期690-694,共5页 Chinese Journal of Biotechnology
基金 上海市科委重点科技攻关项目(No.053919116)资助~~
关键词 猪流感病毒 抗原表位 新型疫苗 swine influenza virus, antigen epitopes, new vaccine
分类号 Q78 [生物学—分子生物学] S852.5 [农业科学—基础兽医学]
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参考文献15

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二级参考文献26

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共引文献27

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二级引证文献29

生物工程学报
2008年 第4期

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