畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (2): 331-339.doi: 10.11843/j.issn.0366-6964.2016.02.016

• 预防兽医 • 上一篇    下一篇

禽呼肠孤病毒强毒株和弱毒疫苗株感染Vero细胞的蛋白质组学研究

黄莉,谢芝勋*,谢丽基,邓显文,谢志勤,范晴,罗思思,黄娇玲,曾婷婷,张艳芳,王盛   

  1. (广西壮族自治区兽医研究所 广西畜禽疫苗新技术重点实验室,南宁 530001)
  • 收稿日期:2015-07-25 出版日期:2016-02-23 发布日期:2016-02-23
  • 通讯作者: 谢芝勋(1963-),研究员,主要从事畜禽传染病分子生物学研究,E-mail: xiezhixun@126.com
  • 作者简介:黄莉(1978-),女,广西南宁人,博士,主要从事禽病防治与病原分子生物学研究,Tel: 0771-3120371, E-mail: lhuang405@126.com
  • 基金资助:

    国家自然科学基金(31160512);广西自然科学基金(2014GXNSFCA118006);广西特聘专家专项(2011B020);广西水产畜牧兽医局科技项目(桂渔牧科201452003)

Proteomic Analysis of Vero Cells Infected by Avian Reovirus Virulent Strain and Attenuated Vaccine Strain

HUANG Li,XIE Zhi-xun*,XIE Li-ji,DENG Xian-wen,XIE Zhi-qin,FAN Qing,LUO Si-si,HUANG Jiao-ling,ZENG Ting-ting,ZHANG Yan-fang,WANG Sheng   

  1. (Guangxi Key Laboratory of Animal Vaccines and New Technology,Guangxi Veterinary Research Institute,Nanning 530001,China)
  • Received:2015-07-25 Online:2016-02-23 Published:2016-02-23

摘要:

为研究在禽呼肠孤病毒(ARV)不同毒力毒株感染后不同时间点宿主细胞中蛋白质组变化,运用双向凝胶电泳技术和质谱鉴定的蛋白质组学方法,对ARV 强毒株S1133、弱毒疫苗株aS1133感染Vero细胞和对照细胞在不同时间点的蛋白质样品进行检测。鉴定结果表明,共鉴定出44个差异表达蛋白质,这些蛋白质包括α-烯醇化酶(α-enolase)、过氧化物酶(Prx-4)、hnRNPs、微管蛋白(Tubulin)、蛋白磷酸酶、转录延伸因子等。 GO软件分析显示这些差异蛋白质主要参与细胞信号、细胞骨架组成、能量代谢、核酸代谢、蛋白质折叠、细胞增殖及凋亡等生物学功能。荧光定量RT-PCR验证表明,hnRNP和Prx-4在S1133感染细胞中表达水平持续上调,而Tubulin 和α-enolase 仅分别在48和24 h时表达上调,与双向凝胶电泳的结果一致。以上试验数据揭示了ARV不同毒力毒株感染后细胞中蛋白质表达的差异变化,有助于进一步阐明ARV和宿主之间的相互作用。

Abstract:

To discover protein expression changes in infected Vero cells with avian reovirus (ARV) virulent strain S1133 and attenuated vaccine strain aS1133,respectively,two dimensional gel electrophoresis (2-DE) and mass spectroscopy (MS) technologies were utilized to separate and identify the total proteins of Vero cells inoculated with ARV S1133 or aS1133 as well as control cells at desired time points post infection.The identification results showed that total 44 differently expressed protein dots were identified,including α-enolase,Prx-4,hnRNPs,tubulin,protein phosphatase,transcription elongation factor.These differentially expressed proteins were involved in various biological functions,including signaling transduction,cytoskeleton,metabolism,protein disfolding,cell proliferation and apoptosis.The results of quantitative RT-PCR validated the mRNA expression of α-enolase,Prx-4,hnRNPs and tubulin were in agreement with that of proteomic analysis.The present data highlighted protein expression differences in infected cells with ARV virulent strain and attenuated vaccine strain,which would contribute to further elucidate the interaction between ARV and hosts.

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