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浙江大学学报（农业与生命科学版）
浙江大学学报（农业与生命科学版）  2019, Vol. 45 Issue (4): 506-511    DOI: 10.3785/j.issn.1008-9209.2018.08.211
动物科学与动物医学     
传染性法氏囊病病毒结构的冷冻电镜初步分析
苗苗1,2,3(),徐彩煌1,2,3,黄子惠1,2,3,张小东4,张兴1,2,3(),吴永平4()
1. 浙江大学医学院生物物理系，杭州 310058
2. 浙江大学医学院附属邵逸夫医院病理科，杭州 310058
3. 浙江大学冷冻电镜中心，杭州 310058
4. 浙江农林大学动物科技学院，杭州 311300
Preliminary structural analysis of infectious bursal disease virus by cryo-electron microscopy
Miao MIAO1,2,3(),Caihuang XU1,2,3,Zihui HUANG1,2,3,Xiaodong ZHANG4,Xing ZHANG1,2,3(),Yongping WU4()
1. Department of Biophysics, Zhejiang University School of Medicine, Hangzhou 310058, China
2. Department of Pathology of Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310058, China
3. The Center of Cryo-Electron Microscopy, Zhejiang University, Hangzhou 310058, China
4. College of Animal Science and Technology, Zhejiang A & F University, Hangzhou 311300, China
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摘要：

为研究传染性法氏囊病病毒（infectious bursal disease virus, IBDV）的结构和基因组在其衣壳内部的包装信息，通过IBDV感染DF-1细胞并分离纯化得到完整的IBDV颗粒，利用单颗粒冷冻电镜技术获得了近生理条件下的IBDV在≈6.6 ?分辨率下的三维结构。结果显示：IBDV病毒衣壳是由260个VP2三聚体蛋白组成的单层衣壳，三角剖分函数T＝13；在IBDV衣壳内部观察不到dsRNA基因组和RNA聚合酶的电子密度，这与其他dsRNA病毒如胞质型多角体病毒（cytoplasmic polyhedrosis virus, CPV）、蓝舌病毒（bluetongue virus, BTV）、轮状病毒（rotavirus, RV）明显不同。该结果暗示，IBDV可能采用了与其他dsRNA病毒不同的方式进行基因组组装、转录和复制，这为进一步揭示IBDV基因组的转录、复制和组装的分子机制及IBDV的起源和进化提供了结构学依据。
关键词： 冷冻电镜;  传染性法氏囊病病毒;  dsRNA基因组结构    
Abstract:

In order to investigate the structure of infectious bursal disease virus (IBDV) and how the double-stranded RNA (dsRNA) segments of IBDV are organized inside its capsid and how the genome segments are transcribed and assembled, we first obtained IBDV particles by infecting DF-1 cell with IBDV and purified the virus particles; we then determined the three-dimensional structure of IBDV using a single-particle cryo-electron microscopy (cryo-EM) at ≈6.6 ? resolution. The results showed that IBDV possessed a T＝13 single shell with 260 VP2 trimers. Unexpectedly no characteristic densities of dsRNA and RNA-dependent RNA polymerase were observed inside the IBDV capsid, which was in contrast to the structures of cytoplasmic polyhedrosis virus (CPV), bluetongue virus (BTV) and rotavirus (RV). These facts implicate that IBDV might adopt different mechanisms to organize, transcribe and assemble its genome segments, and that IBDV might originate and evolve differently from other dsRNA viruses.
Key words: cryo-electron microscopy    infectious bursal disease virus    organization of dsRNA genome
收稿日期: 2018-08-21 出版日期: 2019-09-17
CLC:  S 852.65  
基金资助: 国家重点研发计划(2017YFA0504803);中央高校基本科研业务费专项资金(2018XZZX001-13)
通讯作者: 张兴,吴永平     E-mail: 21618556@zju.edu.cn;xzhang1999@zju.edu.cn;wypzxd2002@163.com;xzhang1999@zju.edu.cn
作者简介: 苗苗（https://orcid.org/0000-0001-6634-8032），E-mail：21618556@zju.edu.cn
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引用本文:

苗苗,徐彩煌,黄子惠,张小东,张兴,吴永平. 传染性法氏囊病病毒结构的冷冻电镜初步分析[J]. 浙江大学学报（农业与生命科学版）, 2019, 45(4): 506-511.

Miao MIAO,Caihuang XU,Zihui HUANG,Xiaodong ZHANG,Xing ZHANG,Yongping WU. Preliminary structural analysis of infectious bursal disease virus by cryo-electron microscopy. Journal of Zhejiang University (Agriculture and Life Sciences), 2019, 45(4): 506-511.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2018.08.211    或    http://www.zjujournals.com/agr/CN/Y2019/V45/I4/506

图1  间接免疫荧光试验检测纯化的IBDV病毒的感染性
图2   IBDV三维重构
图3   IBDV与其他dsRNA病毒内部结构对比

病毒

Virus

衣壳内直径

Inner diameter/nm

衣壳内部体积

Internal volume/nm3

基因组总长（片段数）

Genome length/kbp

（number of segments）

基因组密度

Genome packing

density/（bp/nm3）

传染性法氏囊病病毒

IBDV

 ≈54 ≈82 400 6.0(2) ≈0.072

胞质型多角体病毒

CPV

 ≈52 ≈73 600 24.7(10) ≈0.336

蓝舌病毒

BTV

 ≈48 ≈57 900 19.0(10) ≈0.328

轮状病毒

RV

 ≈47 ≈54 300 18.0(11) ≈0.331
表1  IBDV与其他dsRNA病毒的比较
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