Genomic and Proteomic Analysis of 39 Structural Proteins of Shrimp White Spot Syndrome Virus

39個白點症病毒結構性蛋白質之基因體學與蛋白質體學研究

蔡志明(Jyh-Ming Tsai) ； 王涵青(Han-Ching Wang) ； 羅竹芳(Chu-Fang Lo) ； 郭光雄(Guang-Hsiung Kou)

《Acta Zoologica Taiwanica》 14卷2期 (2004/10) Pp. 99-100

https://doi.org/10.6576/AZT.2004.14.(2).5

摘要

白點症病毒(WSSV)爲嚴重危害世界養蝦產業之重要病毒性病原體，本研究自人工感染螯蝦之血淋巴中純化出之病毒顆粒以梯度丙醯硫胺膠體電泳分離其結構性蛋白質，經胰蛋白酶水解後利用液相層析串聯式質譜儀(LC-nanoESI-MS/MS)進行胺基酸定序。將得到的序列以NCBInr資料庫進行比對，共辨識出33個WSSV結構性蛋白質基因，其中20個爲首次被發表，加上無法在此被辨識之6個已知結構性蛋白質，目前WSSV病毒顆粒至少由39個結構性蛋白質所組成。其結構性蛋白質基因之轉錄表現係利用微陣列分析，多集中於感染晚期(12h.p.i.)才開始表現；依其基因表現趨勢進行叢集分析，將WSSV結構性蛋白質基因分爲四群，同群之基因於病毒感染週期中扮演相似角色。

關鍵字

白點症病毒WSSV ； 液相層析串聯式質譜儀LC-nanoESI-MS/MS ； 結構性蛋白 ； 微陣列分析

並列摘要

White spot syndrome virus (WSSV) is one of the most devastating shrimp pathogens. In this study, WSSV virions were purified from the hemolymph of experimentally infected crayfish Procambarus clarkii and their proteins were separated by 8-18% gradient SDS-PAGE to give a protein profile. We identified 33 WSSV structural genes by LC-nanoESI-MS/MS using a Q-TOF mass spectrometer, twenty of which are reported here for the first time. Since there were six other known WSSV structural proteins that could not be identified from the SDS-PAGE bands, there must therefore be a total of at least39 (33+6) WSSV structural protein genes. Microarray analysis showed that most gene expression started at late stage of infection. Based on similarities in their time course expression patterns, a clustering algorithm was used to group the WSSV structural genes into four clusters. Genes that putatively had common/similar roles in the viral infection cycle tended to appear in the same cluster.

並列關鍵字

white spot syndrome virus WSSV ； LC-nanoESI-MS/MS ； structural protein ； microarray analysis