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非洲马瘟病毒4型在Vero细胞中诱导的多肽特性及其在蛋白质免疫印迹中的反应性

Characterization of African horsesickness virus serotype 4-induced polypeptides in Vero cells and their reactivity in Western immunoblotting.

作者信息

Laviada M D, Arias M, Sánchez-Vizcaíno J M

机构信息

Departamento de Sanidad Animal, CIT-INIA, Madrid, Spain.

出版信息

J Gen Virol. 1993 Jan;74 ( Pt 1):81-7. doi: 10.1099/0022-1317-74-1-81.

Abstract

The structural and non-structural proteins induced by African horsesickness virus serotype 4 (AHSV-4) in infected Vero cells were analysed by SDS-PAGE. Twenty-two virus-induced polypeptides were detected in infected cells by comparison with the polypeptides of mock-infected cells, of which four major (VP2, VP3, VP5 and VP7) and three minor (VP1, VP4 and VP6) structural proteins and four non-structural proteins (P58, P48, P21 and P20) were shown to be virus-coded, as deduced from electrophoretic and antigenic studies of purified virions and infected cells. The proteins that elicit the major antibody responses both in vaccinated and naturally or experimentally infected horses were shown to be three structural proteins, VP2, VP5 and VP7, and the four major non-structural proteins, P58, P48, P21 and P20, as deduced by radioimmunoprecipitation and immunoblotting assays. The cross-reactivity between AHSV-4 and sera obtained from horses experimentally infected with seven other serotypes was also determined. The results showed that VP5, VP7, P48, P21 and P20 are conserved and can be used to diagnose the infection of any of these eight serotypes.

摘要

采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析了非洲马瘟病毒4型(AHSV-4)感染的非洲绿猴肾(Vero)细胞中诱导产生的结构蛋白和非结构蛋白。与 mock 感染细胞的多肽相比,在感染细胞中检测到22种病毒诱导的多肽,其中4种主要结构蛋白(VP2、VP3、VP5和VP7)、3种次要结构蛋白(VP1、VP4和VP6)以及4种非结构蛋白(P58、P48、P21和P20)经纯化病毒粒子和感染细胞的电泳及抗原研究推断为病毒编码。通过放射免疫沉淀和免疫印迹分析推断,在接种疫苗以及自然或实验感染马匹中引发主要抗体反应的蛋白为3种结构蛋白VP2、VP5和VP7以及4种主要非结构蛋白P58、P48、P21和P20。还测定了AHSV-4与从实验感染其他7种血清型的马匹获得的血清之间的交叉反应性。结果表明,VP5、VP7、P48、P21和P20是保守的,可用于诊断这8种血清型中任何一种的感染。

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