Grubman M J, Appleton J A, Letchworth G J
Virology. 1983 Dec;131(2):355-66. doi: 10.1016/0042-6822(83)90503-2.
Double-stranded (ds) RNA was isolated from either partially purified bluetongue virus (BTV) type 17 or BTV-17 infected cells. The RNA was denatured with either methylmercury hydroxide or dimethyl sulfoxide and translated in a reticulocyte cell-free system. The translation products were found to be similar to BTV-17 specific polypeptides in infected cells when analyzed by polyacrylamide gel electrophoresis and immunoprecipitation with BTV-17 specific polyclonal mouse ascitic fluid. The 10 dsRNA BTV-17 genome segments were resolved by preparative gel electrophoresis and the coding assignment of each segment was determined by translation in a cell-free system. Immunoprecipitation of VP3 from infected cell lysates by a type-specific neutralizing monoclonal antibody identified this polypeptide, which is coded for by genome segment 2, as being involved in virus neutralization. Immunoprecipitation of in vitro translation products directed by total BTV-17 RNA with a monoclonal antibody that reacts with all the serotypes of BTV and some other orbiviruses and precipitates VP7 directly demonstrated that gene segment 8 codes for this intergroup-specific polypeptide.
双链（ds）RNA 从部分纯化的 17 型蓝舌病毒（BTV）或 BTV - 17 感染的细胞中分离得到。该 RNA 用氢氧化甲基汞或二甲基亚砜变性，并在网织红细胞无细胞体系中进行翻译。当通过聚丙烯酰胺凝胶电泳以及用 BTV - 17 特异性多克隆小鼠腹水进行免疫沉淀分析时，发现翻译产物与感染细胞中的 BTV - 17 特异性多肽相似。通过制备性凝胶电泳分离出 10 个 BTV - 17 基因组双链 RNA 片段，并通过在无细胞体系中的翻译确定每个片段的编码任务。用型特异性中和单克隆抗体对感染细胞裂解物中的 VP3 进行免疫沉淀，鉴定出该多肽（由基因组片段 2 编码）参与病毒中和。用一种能与所有 BTV 血清型及其他一些环状病毒发生反应并直接沉淀 VP7 的单克隆抗体对由总 BTV - 17 RNA 指导的体外翻译产物进行免疫沉淀，直接证明基因片段 8 编码这种组间特异性多肽。
