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犬冠状病毒基因重组抗原间接ELISA检测方法的研究 被引量:3
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作者 乔军 夏咸柱 +3 位作者 胡桂学 孙贺廷 杨松涛 黄耕 《吉林农业大学学报》 CAS CSCD 北大核心 2004年第3期330-333,共4页
以在E coli高效表达的犬冠状病毒核蛋白为抗原,用辣根过氧化物酶(HRP)标记的兔抗犬IgG为二抗,建立了检测犬冠状病毒抗体的间接ELISA方法。经方阵滴定确定出最佳反应条件为1μg 孔纯化的E coli表达的重组N蛋白抗原包被酶标板,用10%的兔... 以在E coli高效表达的犬冠状病毒核蛋白为抗原,用辣根过氧化物酶(HRP)标记的兔抗犬IgG为二抗,建立了检测犬冠状病毒抗体的间接ELISA方法。经方阵滴定确定出最佳反应条件为1μg 孔纯化的E coli表达的重组N蛋白抗原包被酶标板,用10%的兔血清进行封闭,以正常E coli裂解上清液稀释待检血清。结果表明:应用重组N蛋白作为诊断用抗原具有特异性强、稳定性高、成本较低等特点。 展开更多
关键词 犬冠状病毒 基因重组核蛋白 间接ELISA
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Gene cloning and prokaryotic expression of recombinant flagellin A from Vibrio parahaemolyticus 被引量:2
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作者 袁野 王秀利 +3 位作者 郭设平 刘洋 葛辉 仇雪梅 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2010年第6期1254-1260,共7页
The Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals. Bacteria flagellins play an important role during infection and induction of the host immune response. Thus, flagellin prote... The Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals. Bacteria flagellins play an important role during infection and induction of the host immune response. Thus, flagellin proteins are an ideal target for vaccines. We amplified the complete flagellin subunit gene (tTaA) from V. parahaemolyticus ATCC 17802. We then cloned and expressed the gene into Escherichia coli BL21 (DE3) cells. The gene coded for a protein that was 62.78 kDa. We purified and characterized the protein using Ni-NTA affinity chromatography and Anti-His antibody Western blotting, respectively. Our results provide a basis for further studies into the utility of the FlaA protein as a vaccine candidate against infection by Vibrio parahaemolyticus. In addition, the purified FlaA protein can he used for further functional and structural studies. 展开更多
关键词 Vibrio parahaemolyticus flagellin subunit gene (flail) CLONING prokaryotic expression characterization
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