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虎纹蛙病毒可感染斑马鱼细胞,用于通过蛋白质组学方法研究上调或下调的基因。

Tiger frog virus can infect zebrafish cells for studying up- or down-regulated genes by proteomics approach.

作者信息

Luo Yongwen, Weng Shaoping, Wang Qing, Shi Xiujuan, Dong Chuanfu, Lu Qingxia, Yu Xiaoqiang, He Jianguo

机构信息

State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen (Zhongshan) University, Guangzhou, 510275, China.

出版信息

Virus Res. 2009 Sep;144(1-2):171-9. doi: 10.1016/j.virusres.2009.04.016. Epub 2009 May 4.

Abstract

Tiger frog virus (TFV), a member of the iridovirus family, causes high mortality of cultured tiger frog tadpoles in southern China. To better understand TFV infection and its interaction with host cells, zebrafish embryonic fibroblast (ZF4) cells, a stable polyploid cell line with most clear genetic map, was used for our present study. Our results showed that TFV caused typical lytic plaque forming cytopathic effect (CPE) and that various stages of viral proliferation were observed using electron microscopy and indirect immunofluorescence assay. Two-dimensional electrophoresis also showed that some cellular proteins were differentially expressed in the ZF4 cells infected with TFV. A total of 10 proteins were identified using the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) technique, including 7 that were up-regulated and 3 that were down-regulated after infection. Among the 10 identified proteins, alterations in Hsp90 and alpha-tubulin expression were further confirmed by Western blot analysis. Furthermore, reorganization of microtubules was also observed in TFV-infected cells and can probably be attributed to the overexpression of translationally controlled tumor protein. The present study is the first attempt to reveal cellular responses to TFV infection by proteomics. The results suggest that the ZF4 cell line could be used as a model to study TFV infection.

摘要

虎纹蛙病毒(TFV)是虹彩病毒科的成员,在中国南方导致养殖的虎纹蛙蝌蚪高死亡率。为了更好地了解TFV感染及其与宿主细胞的相互作用,本研究使用了斑马鱼胚胎成纤维细胞(ZF4),这是一种具有最清晰遗传图谱的稳定多倍体细胞系。我们的结果表明,TFV引起典型的溶菌斑形成细胞病变效应(CPE),并且使用电子显微镜和间接免疫荧光测定法观察到了病毒增殖的各个阶段。二维电泳还表明,在感染TFV的ZF4细胞中一些细胞蛋白存在差异表达。使用基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)技术共鉴定出10种蛋白质,其中7种在感染后上调,3种下调。在鉴定出的10种蛋白质中,通过蛋白质印迹分析进一步证实了Hsp90和α-微管蛋白表达的变化。此外,在感染TFV的细胞中还观察到微管的重组,这可能归因于翻译控制肿瘤蛋白的过表达。本研究是首次尝试通过蛋白质组学揭示细胞对TFV感染的反应。结果表明,ZF4细胞系可作为研究TFV感染的模型。

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