Lanser J A, Howard C R
J Gen Virol. 1980 Feb;46(2):349-61. doi: 10.1099/0022-1317-46-2-349.
The Massachusetts strain of avian infectious bronchitis virus was purified from embryonated hens' eggs. Four major species of apparent mol. wt. 90 000, 52 000, 29 000 and 26 000 were resolved by SDS-polyacrylamide gel electrophoresis. Omission of reducing agent failed to resolve the 29 000 mol. wt. component. Labelling of acrylamide gels with 125I-concanavalin A indicated that polypeptides of mol. wt. 90 000, 29 000 and 26 000 were glycosylated and, in the absence of reducing agent, that the 29 000 species migrated as a dimer in the 5000 mol. wt. region. Purified IBV radio-iodinated with Bolton and Hunter reagent, which banded as a single peak of radioactivity in Metrizamide gradients, was found to contain bands of radioactivity when analysed by SDS-PAGE, corresponding to the polypeptides of mol. wt. 90 000, 52 000 and 29 000 resolved in stained gels. Disruption of IBV particles in Triton X-100 released two subviral particles, a 16 nm spike which comprised polypeptides of 90 000, 52 000 and 29 000 mol. wt. and another denser spherical particle of 25 to 45 nm which contained RNA and the 52 000 and 26 000 polypeptides.
从感染的母鸡胚胎蛋中提纯出了马萨诸塞州禽传染性支气管炎病毒株。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳解析出了四种表观分子量分别为90000、52000、29000和26000的主要蛋白条带。去除还原剂后未能解析出分子量为29000的组分。用125I-伴刀豆球蛋白A对丙烯酰胺凝胶进行标记表明,分子量为90000、29000和26000的多肽是糖基化的,并且在没有还原剂的情况下,分子量为29000的蛋白条带在5000分子量区域以二聚体形式迁移。用博尔顿-亨特试剂进行放射性碘化处理的纯化传染性支气管炎病毒(IBV),在甲泛影酰胺梯度中呈单一放射性峰,经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析发现含有放射性条带,对应于染色凝胶中解析出的分子量为90000、52000和29000的多肽。在Triton X-100中破坏传染性支气管炎病毒粒子释放出两种亚病毒粒子,一种16纳米的纤突,由分子量为90000、52000和29000的多肽组成,另一种密度更大的25至45纳米球形粒子,含有RNA以及分子量为52000和26000的多肽。