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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Nucleocapsid protein N of Lelystad virus: expression by recombinant baculovirus, immunological properties, and suitability for detection of serum antibodies.
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Nucleocapsid protein N of Lelystad virus: expression by recombinant baculovirus, immunological properties, and suitability for detection of serum antibodies.

机译：Lelystad病毒的核衣壳蛋白N：重组杆状病毒的表达，免疫学性质以及对检测血清抗体的适用性。

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摘要

The ORF7 gene, encoding the nucleocapsid protein N of Lelystad virus (LV), was inserted downstream of the P10 promoter into Autographa californica nuclear polyhedrosis virus (baculovirus). The resulting recombinant baculovirus, designated bac-ORF7, expressed a 15-kDa protein in insect cells. This protein was similar in size to the N protein expressed by LV in CL2621 cells when it was analyzed on sodium dodecyl sulfate-polyacrylamide gels. The N protein expressed by bac-ORF7 was immunoprecipitated with anti-ORF7 was immunoprecipitated with anti-ORF7 peptide serum, porcine convalescent-phase anti-LV serum, and N protein-specific monoclonal antibodies, indicating that this N protein had retained its native antigenic structure. The recombinant N protein was immunogenic in pigs, and the porcine antibodies raised against this protein recognized LV in an immunoperoxidase monolayer assay. However, pigs vaccinated twice with approximately 20 micrograms of N protein were not protected against a challenge with 10(5) 50% tissue culture infective doses of LV. Experimental and field sera directed against various European and North American isolates reacted with the N protein expressed by bac-ORF7 in a blocking enzyme-linked immunosorbent assay. Therefore, the recombinant N protein may be useful for developing diagnostic assays for the detection of serum antibodies directed against different isolates of LV.
机译：将编码Lelystad病毒（LV）的核衣壳蛋白N的ORF7基因插入到P10启动子的下游，插入到加利福尼亚州的Autographa californica核多角体病毒（杆状病毒）中。所得的重组杆状病毒，称为bac-ORF7，在昆虫细胞中表达15 kDa的蛋白质。当在十二烷基硫酸钠-聚丙烯酰胺凝胶上分析该蛋白质时，其大小与LV在CL2621细胞中表达的N蛋白相似。 bac-ORF7表达的N蛋白用抗ORF7免疫沉淀，用抗ORF7肽血清，猪恢复期抗LV血清和N蛋白特异性单克隆抗体免疫沉淀，表明该N蛋白保留了其天然抗原性结构体。重组N蛋白在猪中具有免疫原性，针对该蛋白的猪抗体在免疫过氧化物酶单层测定中可识别LV。但是，用约20微克N蛋白接种两次的猪没有受到10（5）50％组织培养感染剂量的LV的攻击。在封闭的酶联免疫吸附试验中，针对各种欧洲和北美分离株的实验血清和野外血清与bac-ORF7表达的N蛋白发生了反应。因此，重组N蛋白可用于开发诊断检测以检测针对LV不同分离株的血清抗体。

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