基于iTRAQ技术伪狂犬病毒感染宿主细胞差异表达蛋白质组分析

发布时间：2018-09-17 18:28

【摘要】：为了研究伪狂犬病毒(pseudorabies virus,PRV)感染宿主细胞后宿主蛋白的差异表达情况,寻找PRV与细胞相互作用的重要分子,本试验将PRV接种PK-15细胞,运用同位素标记(isobaric tags for relative and absolute quantitation,iTRAQ)结合色谱/串联质谱联用技术,开展病毒感染细胞后差异表达蛋白分析,并通过qPCR对其数据进行验证。以未接毒细胞为对照,通过数据库检索,分析出3 062个蛋白,共鉴定到27个差异表达蛋白,其中21个蛋白下调,6个蛋白上调。这些差异蛋白主要参与代谢过程、生物调控、应激反应以及定位等生物学过程,具有结合活性、催化活性、转录活性等分子功能。NFκB1、PIK3R2、HSPA1L等差异蛋白在多条通路中发挥作用。POP1、UTP15和ND4的表达变化情况与iTRAQ结果变化趋势一致,证明了该数据的可靠性。本试验研究结果为进一步研究PRV致病机制及宿主免疫应答提供理论依据。
[Abstract]:In order to study the differential expression of host proteins after pseudorabies virus (pseudorabies virus,PRV) infection in host cells and to find the important molecules of interaction between PRV and cells, PRV was inoculated into PK-15 cells. The differential expression protein analysis of virus infected cells was carried out by using isotope labeled (isobaric tags for relative and absolute quantitation,iTRAQ and chromatography-tandem mass spectrometry (GC-MS), and its data were verified by qPCR. Using uninfected cells as control, 3 062 proteins were identified and 27 differentially expressed proteins were identified, of which 21 proteins were down-regulated and 6 proteins up-regulated. These differential proteins are mainly involved in metabolic processes, biological regulation, stress response and localization, and have binding activity and catalytic activity. Transcriptional activity and other molecular functions, such as NF 魏 B 1, PIK3R 2, HSPA1L, play a role in many pathways. The changes of the expression of POP 1 UTP15 and ND4 are consistent with the change trend of iTRAQ results, which proves the reliability of the data. The results of this study provide theoretical basis for further study on the pathogenesis of PRV and host immune response.
【作者单位】：西北农林科技大学动物医学院;中国农业科学院上海兽医研究所;
【基金】：上海市自然科学基金资助项目(14ZR1448900) 上海市科技兴农重点攻关资助项目(沪农科攻字(2015)第1-7号)
【分类号】：S852.65

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