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风疹病毒与仓鼠潜伏逆转录病毒杂交体的结构基因产物及形态发生

Structural gene products and morphogenesis of a hybrid between rubella virus and a hamster latent retrovirus.

作者信息

Urade M, Mima T, Yamamoto N

机构信息

First Department of Oral and Maxillofacial Surgery, Osaka University Faculty of Dentistry, Japan.

出版信息

Res Virol. 1993 Mar-Apr;144(2):129-39. doi: 10.1016/s0923-2516(06)80021-x.

Abstract

We studied the structural gene products and morphogenesis of the hybrid between rubella virus (RV) and a latent retrovirus (R-type virus or R virus) of a baby hamster kidney cell line, BHK21/WI-2. In electron microscopic analysis, the rubella virion measures 80 nm in diameter and has a round nucleoid form with an electron-lucid centre, while R-type virus is 110 nm in diameter and has a round nucleoid with radial spokes. The type 2 hybrid (H2) virion is pleomorphic, ranging from 85 to 110 nm in diameter, and has a nucleoid structure similar to R virus, with short spokes. Indirect immunofluorescence staining of H2-infected cells with polyclonal and monoclonal antibodies (mAb) against the RV structural proteins demonstrated expression of the homologous RV gene products. SDS-PAGE of 35S-methionine-labelled proteins in RV- and H2-infected cells revealed that they both produce four major immunoprecipitable proteins, three of which (E2a, E2b and C) are products of the two genetic homologous loci (e2 and c). In nonhomologous regions, RV-infected cells exhibited E1 protein, while H2-infected cells produced the Env protein of R virus. Western immunoblotting analysis of RV and H2 viral proteins with mAb confirmed that H2 virions carry rubella viral E2 and C proteins and that E1 protein is carried by RV but not H2 virion. These results explained the absence of serological cross-neutralization of haemagglutination and plaque-forming ability of RV and H2 viruses.

摘要

我们研究了风疹病毒(RV)与幼仓鼠肾细胞系BHK21/WI-2的潜伏逆转录病毒(R型病毒或R病毒)之间杂交体的结构基因产物和形态发生。在电子显微镜分析中,风疹病毒粒子直径为80nm,具有圆形核样体形式,中心电子透明,而R型病毒直径为110nm,具有带放射状辐条的圆形核样体。2型杂交体(H2)病毒粒子多形性,直径范围为85至110nm,具有与R病毒相似的核样体结构,辐条较短。用针对RV结构蛋白的多克隆和单克隆抗体(mAb)对H2感染细胞进行间接免疫荧光染色,证明了同源RV基因产物的表达。对RV和H2感染细胞中35S-甲硫氨酸标记蛋白进行SDS-PAGE分析表明,它们都产生四种主要的可免疫沉淀蛋白,其中三种(E2a、E2b和C)是两个基因同源位点(e2和c)的产物。在非同源区域,RV感染细胞表现出E1蛋白,而H2感染细胞产生R病毒的Env蛋白。用mAb对RV和H2病毒蛋白进行Western免疫印迹分析证实,H2病毒粒子携带风疹病毒E2和C蛋白,E1蛋白由RV携带而不是H2病毒粒子。这些结果解释了RV和H2病毒在血凝和蚀斑形成能力方面缺乏血清学交叉中和作用的原因。

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