高致病性猪繁殖与呼吸综合征病毒感染MARC-145细胞的差异蛋白

doi:10.3864/j.issn.0578-1752.2012.04.020

摘要/Abstract

摘要： 【目的】研究高致病性猪繁殖与呼吸综合征病毒（PRRSV）感染MARC-145细胞生长相关蛋白表达量的变化，明确差异蛋白的功能信息，为深入研究PRRSV病毒复制和致病机理奠定基础。【方法】采用差异蛋白质组学和Western blotting技术，通过建立双向凝胶电泳，对高致病性PRRSV感染MARC-145细胞后表达量差异蛋白质进行了胶内酶解、质谱分析鉴定及数据库检索，明确差异蛋白质的相关信息。【结果】得到7个表达稳定、重复性好的蛋白点，其中有5个蛋白点找到匹配蛋白，分别为热休克蛋白70、硫氧还蛋白、S100钙结合蛋白A6、3-磷酸甘油脱氢酶和内参肌动蛋白，其余2个蛋白点为未知蛋白。在MARC-145细胞感染PRRSV后，热休克蛋白70和S100钙结合蛋白A6表现为下调，而硫氧还蛋白、3-磷酸甘油脱氢酶和内参肌动蛋白表现为上调。功能预测分析发现，5个蛋白与MARC-145细胞抗应激、抗氧化、生长、凋亡和信号转导等作用息息相关。【结论】高致病性PRRSV感染MARC-145细胞后，能诱导细胞部分功能蛋白表达量的增加或减少。

关键词: 猪繁殖与呼吸综合征病毒, MARC-145细胞, 蛋白质组学

Abstract: 【Objective】 The objective of this study is to analyze the protein expression changes of the MARC-145 which was infected with highly pathogenic PRRSV. Proteomic techniques were used to establish the protein expression profiles. The study provides the global information of proteomic alteration of MARC-145 cells in the presence of intact PRRSV and the clues for further understanding of the mechanism of interaction. 【Method】 The differentially expressed proteins between the MARC-145 cells that were infected with highly pathogenic PRRSV and the normal MARC-145 cells were profiled and identified by two dimensional electrophoresis and Western Blotting. Differential proteinaceous information was identified by mass chromatographic analysis and retrieval. 【Result】The results showed that there were seven differentially expressed proteins. Five proteins were identified successfully which were heat-shock 70-kD protein, thioredoxin, S100 calcium-binding protein A6, glyceraldehyde-3-phosphate dehydrogenase, and beta-actin, respectively. Among them, the heat-shock 70-kD protein and S100 calcium-binding protein A6 were down-regulated proteins; thioredoxin, glyceraldehyde-3-phosphate dehydrogenase, and beta-actin were up-regulated proteins. The function of the five proteins were closely associated with the growth, apoptosis, santioxidant and ignal transduction of MARC-145 cells. 【Conclusion】The quantity of functional proteins of the MARC-145 was changed which were infected by highly pathogenic PRRSV.

Key words: porcine productive and respiratory syndrome virus, MARC-145 cells, proteomics

周伦江, 王隆柏, 方勤美, 吴学敏, 车勇良, 陈如敬, 魏宏, 庄向生. 高致病性猪繁殖与呼吸综合征病毒感染MARC-145细胞的差异蛋白[J]. 中国农业科学, 2012, 45(4): 786-793.

ZHOU Lun-Jiang, WANG Long-Bai, FANG Qin-Mei, WU Xue-Min, CHE Yong-Liang, CHEN Ru-Jing, WEI Hong, ZHUANG Xiang-Sheng. Differential Proteome Analysis of the MARC-145 Cell Infected with Highly Pathogenic PRRSV[J]. Scientia Agricultura Sinica, 2012, 45(4): 786-793.

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