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利用电子冷冻显微镜技术解析螨传黑醋栗退化线虫传多面体病毒的结构

Structure of the mite-transmitted Blackcurrant reversion nepovirus using electron cryo-microscopy.

作者信息

Seitsonen Jani J T, Susi Petri, Lemmetty Anne, Butcher Sarah J

机构信息

Institute of Biotechnology, P.O. Box 65 (Viikinkaari 1), FIN-00014 University of Helsinki, Finland.

出版信息

Virology. 2008 Aug 15;378(1):162-8. doi: 10.1016/j.virol.2008.05.005. Epub 2008 Jun 16.

Abstract

Blackcurrant reversion nepovirus (BRV; genus Nepovirus) has a single-stranded, bipartite RNA genome surrounded by 60 copies of a single capsid protein (CP). BRV is the most important mite-transmitted viral pathogen of the Ribes species. It is the causal agent of blackcurrant reversion disease. We determined the structure of BRV to 1.7 nm resolution using electron cryo- microscopy (cryoEM) and image reconstruction. The reconstruction reveals a pseudo T=3 viral capsid similar to that of tobacco ringspot virus (TRSV). We modelled the BRV capsid protein to that of TRSV and fitted it into the cryoEM reconstruction. The fit indicated that the extended C-terminus of BRV-CP is located on the capsid surface and the N-terminus on the interior. We generated peptide antibodies to two putatively exposed C-terminal sequences and these reacted with the virus. Hence homology modelling may be useful for defining epitopes for antibody generation for diagnostic testing of BRV in commercial crops.

摘要

黑加仑逆转线状病毒(BRV;线状病毒属)具有单链双分体RNA基因组,由60个单一衣壳蛋白(CP)拷贝包围。BRV是醋栗属植物中最重要的螨传病毒病原体。它是黑加仑逆转病的病原体。我们使用冷冻电子显微镜(cryoEM)和图像重建技术,将BRV的结构分辨率确定为1.7纳米。重建结果显示出一种类似于烟草环斑病毒(TRSV)的伪T=3病毒衣壳。我们将BRV衣壳蛋白建模为TRSV的衣壳蛋白,并将其拟合到冷冻电镜重建中。拟合结果表明,BRV-CP的延伸C末端位于衣壳表面,N末端位于内部。我们针对两个假定暴露的C末端序列产生了肽抗体,这些抗体与病毒发生了反应。因此,同源建模可能有助于确定用于商业作物中BRV诊断检测的抗体产生的表位。

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