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通过二维电泳-质谱联用技术鉴定对虾白斑综合征病毒(WSSV)的结构蛋白

Identification of structural proteins from shrimp white spot syndrome virus (WSSV) by 2DE-MS.

作者信息

Zhang Xiaobo, Huang Canhua, Tang Xuhua, Zhuang Ying, Hew Choy Leong

机构信息

Department of Biological Sciences, National University of Singapore, Singapore.

出版信息

Proteins. 2004 May 1;55(2):229-35. doi: 10.1002/prot.10640.

Abstract

White spot syndrome virus (WSSV) is a major shrimp pathogen that also infects many other species of crustaceans. Its 305-kb double-stranded DNA genome has the capacity to encode 181 presumptive proteins. In an attempt to identify the viral proteins from the 181 theoretical proteins, proteins of the purified WSSV were separated by two-dimensional electrophoresis (2-DE). More than 60 protein spots were revealed, as detected by silver staining, from which 12 viral proteins were identified by mass spectrometry. In total, 25 WSSV proteins, including those reported in one of our earlier studies (Huang et al., Mol Cell Proteomics 2002;1:223-231), were revealed by this proteomic approach, and their corresponding genes were further confirmed by reverse transcription-polymerase chain reaction (RT-PCR). Two of them were characterized to be WSSV envelope proteins using immuno-electron microscopy. Our study showed that the proteomic approach is a powerful method for discovering the viral structural proteins and their corresponding genes.

摘要

白斑综合征病毒(WSSV)是一种主要的虾类病原体,也能感染许多其他种类的甲壳类动物。其305千碱基对的双链DNA基因组能够编码181种推定蛋白。为了从这181种理论蛋白中鉴定出病毒蛋白,通过二维电泳(2-DE)分离纯化的WSSV蛋白。经银染检测,发现了60多个蛋白点,通过质谱鉴定出其中12种病毒蛋白。通过这种蛋白质组学方法总共鉴定出25种WSSV蛋白,包括我们早期一项研究(Huang等人,《分子细胞蛋白质组学》2002年;1:223 - 231)中报道的那些蛋白,并且通过逆转录 - 聚合酶链反应(RT-PCR)进一步确认了它们相应的基因。其中两种蛋白通过免疫电子显微镜被鉴定为WSSV包膜蛋白。我们的研究表明,蛋白质组学方法是发现病毒结构蛋白及其相应基因的有力方法。

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