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Rabies diagnostic reagents prepared from a rabies N gene recombinant expressed in baculovirus.

机译:由在杆状病毒中表达的狂犬病N基因重组体制备的狂犬病诊断试剂。

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摘要

A gene encoding the nucleoprotein (N) of rabies virus was inserted into the genome of the baculovirus Autographa californica nuclear polyhedrosis virus. Recombinant gene expression was controlled by the strong polyhedrin gene promoter. Insect cells (Spodoptera frugiperda) infected by a baculovirus recombinant containing the rabies virus N gene produced abundant amounts of a novel 55-kilodalton protein of a size comparable to that of the rabies virus N protein, as demonstrated by polyacrylamide gel electrophoresis. This new gene product possessed the antigenic and immunogenic properties of native viral N protein, as shown by the ability of the new protein to react in immunoprecipitation and immunofluorescence assays with antirabies antibodies, to serve as a substitute for infectious rabies virus in adsorbing suspensions for diagnostic tests, and to induce high-titered antiserum. The baculovirus expression system provides a safe, convenient, and inexpensive source of rabies virus N protein for the production of both antiserum and adsorbing suspensions for use in rabies diagnoses.
机译:将编码狂犬病病毒核蛋白(N)的基因插入杆状病毒加州苜蓿白纹病核多角体病毒的基因组中。重组基因表达由强多面体基因启动子控制。如通过聚丙烯酰胺凝胶电泳所证实的那样,被含有狂犬病病毒N基因的杆状病毒重组体感染的昆虫细胞(Spodoptera frugiperda)产生了大量的新型55-奇洛顿蛋白,其大小与狂犬病病毒N蛋白的大小相当。这种新的基因产物具有天然病毒N蛋白的抗原和免疫原性,如新蛋白在与抗狂犬病抗体的免疫沉淀和免疫荧光测定中反应的能力所证明,可替代传染性狂犬病毒在吸附悬浮液中进行诊断测试,并诱导高滴度抗血清。杆状病毒表达系统提供了狂犬病病毒N蛋白的安全,方便和廉价来源,可用于生产抗血清和吸附性悬浮液,用于狂犬病诊断。

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