高等学校化学学报

高等学校化学学报 ›› 2011, Vol. 32 ›› Issue (8): 1768.

• 研究论文 • 上一篇    下一篇

中蜂囊状幼虫病毒结构蛋白预测与鉴定

马鸣潇1, 马臣2, 程健1, 谢振声3, 李明1, 费东亮1   

  1. 1.  辽宁医学院实验动物中心, 锦州 121001;
    2.  赤峰市翁牛特旗中蒙医院, 乌丹 024500;
    3.  中国科学院生物物理研究所蛋白质组学平台, 北京 100101
  • 收稿日期:2011-01-06 修回日期:2011-03-18 出版日期:2011-08-10 发布日期:2011-07-19
  • 通讯作者: 马鸣潇 E-mail:lnjzmmx@163.com
  • 基金资助:

    国家自然科学基金(批准号: 30972200)和辽宁省教育厅项目(批准号: 2009A464)资助.

Prediction and Identification of Structural Proteins of Chinese Sacbrood Virus LN Isolate

MA Ming-Xiao1*, MA Chen2, CHENG Jian1, XIE Zhen-Sheng3, LI Ming1, FEI Dong-Liang1   

  1. 1. Department of Laboratory Animal Center Liaoning Medical University, Jinzhou 121001, China;
    2. Wengniuete Country Chinese Traditional Medicine and Mongolian Medicine in Chifeng Town, Wudan 024500, China;
    3. Proteomic Platform,   Institute of Biophysics,   Chinese Academy of Sciences, Beijing  100101, China
  • Received:2011-01-06 Revised:2011-03-18 Online:2011-08-10 Published:2011-07-19
  • Contact: MA Ming-Xiao E-mail:lnjzmmx@163.com
  • Supported by:

    国家自然科学基金(批准号: 30972200)和辽宁省教育厅项目(批准号: 2009A464)资助.

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被引次数

15

摘要： 摘要 本文采用RT-PCR方法和质谱分析对中蜂囊状幼虫病毒辽宁株（CSBV-LN）全基因组进行分子克隆、序列测定、分子生物学特性分析及其结构蛋白预测和鉴定。结果获得CSBV-LN全基因组序列长为8863个核苷酸，编码一个大的开放阅读框，其中包括178nt的5'非编码区的前导序列和142nt的3'非编码区，后面跟着一个poly(A)的尾巴。通过软件分析表明，CSBV-LN蛋白序列，类似于哺乳动物的小RNA病毒蛋白序列，预计存在VP1，VP2， VP3和VP4四个小的结构蛋白，系统分析进化分析表明CSBV，SBV和 DWV起源相同，推测CSBV有着类似DWV的结构蛋白序列5’-VP2 VP4 VP1 VP3-3。基于此，根据PAGE蛋白条带的大小，利用NetPicoRNA软件预测结构蛋白的裂解位点。在此基础上，对纯化后的CSBV进行SDS-PAGE，经SDS-PAGE分离到的4个蛋白进行质谱分析，结果鉴定出VP1、VP3和VP0三个蛋白。

关键词: 中蜂囊状幼虫病毒; , 反转录聚合酶链反应; , 结构蛋白; , 质谱分析

Abstract: Abstract：Chinese sacbrood virus (CSBV) was purified from diseased insects, and its genome was cloned and sequenced. The genomic RNA of CSBV is 8863 nucleotides in length and contains a single large open reading frame encoding a 319.614kDa polyprotein. The coding sequence is flanked by a 178-nucleotide 5' nontranslated leader sequence and a 142-nucleotide 3' nontranslated region, followed a poly(A) tail. Four major structural proteins, VP1,VP2, VP3 and VP4, were estimated, and mapped to the N-terminal section of the polyprotein of which the genes were predicted. Phylogenetic analysis using RdRp of selected picorna-like viruses shows that CSBV/SBV and Deformed Wing Virus (DWV) tend to group together, which possesses an RNA of similar size and gene order. The results predicted the probable size and gene order of CSBV/SBV. Structural proteins, VP1, VP3 and VP0, were identified after four proteins was analysed by mass spectrographic.

Key words: Chinese sacbrood virus(CSBV), RT-PCR, Structural protein; , Mass spectrographic analysis

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