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首页> 外文会议>16th International congress on animal hygiene : animal hygiene, health and welfare as corner stones of sustainable animal production >Expression in Insect Cells and Antigenic Analysis of Recombinant Nucleocapsid Protein of Canine Distemper Virus
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Expression in Insect Cells and Antigenic Analysis of Recombinant Nucleocapsid Protein of Canine Distemper Virus

机译：犬瘟热病毒在昆虫细胞中的表达及重组核衣壳蛋白的抗原分析

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摘要

The nucleocapsid (N)protein gene of an atypical Canine distemper virus (CDV)was cloned into pFastBacHTA donor plasmid.The recombinant donor plasmid pFastBac-N containing N gene was constructed and transformed into competent E.coli DHIOBac cells that were grown on LB plate containing 3 antibiotics.Recombinant Bacmid DNA (rBacmid-N)was obtained and used to transfect insect cells Sf9 with Lipofectamine to produce baculovirus vBacmid-N.The expressed recombinant N protein band of approximate 62 kDa was detected in Western blot.The recombinant vBacmid-N antigen was visualized in infected Sf9 cells in indirect immunofluorescence assay.Purified recombinant N protein was used to establish indirect ELISA for the detection of antibody against CDV.The absorbance values at 450 nm of all CD-positive serum samples were above 0.4 whereas CD-negative serum samples were below 0.05.
机译：将非典型犬瘟热病毒（CDV）的核衣壳（N）蛋白基因克隆到pFastBacHTA供体质粒中，构建了含有N基因的重组供体质粒pFastBac-N，并将其转化为能在LB平板上生长的大肠杆菌DHIOBac细胞含有3种抗生素。获得重组Bacmid DNA（rBacmid-N），并用Lipofectamine转染昆虫细胞Sf9以生产杆状病毒vBacmid-N.Western blot检测到表达的重组N蛋白条带约为62 kDa。间接免疫荧光法在感染的Sf9细胞中可见N抗原，使用纯化的重组N蛋白建立间接ELISA检测CDV抗体，所有CD阳性血清样品在450 nm处的吸光度均高于0.4，而CD-阴性血清样品低于0.05。

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  • 会议地点 Nanjing(CN)
  • 作者

    Bi Zhenwei; Wang Yongshan; Ouyang Wei; Fan HongJie;

  • 作者单位

    Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences/Key Laboratory of Veterinary Biological Engineering and Technology,Ministry of Agriculture/National Center for Engineering Research of Veterinary Bio-products,Nanjing 210014,China;

    Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences/Key Laboratory of Veterinary Biological Engineering and Technology,Ministry of Agriculture/National Center for Engineering Research of Veterinary Bio-products,Nanjing 210014,China;

    Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences/Key Laboratory of Veterinary Biological Engineering and Technology,Ministry of Agriculture/National Center for Engineering Research of Veterinary Bio-products,Nanjing 210014,China;

    College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 兽医公共卫生学（家畜卫生学）;
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