畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (12): 1995-1999.doi: 10.11843/j.issn.0366-6964.2014.12.012

• 预防兽医 • 上一篇    下一篇

猪瘟病毒C蛋白与宿主β-actin相互作用的鉴定

董泓1,李丹2,陈佳宁2,李素2,何文瑞2,冯烁2,贺番2,廖亚金2,孙元2,胡永浩1*   

  1. (1.甘肃农业大学 动物医学院,兰州 730000;2.中国农业科学院哈尔滨兽医研究所 兽医生物技术国家重点实验室,哈尔滨 150001)
  • 收稿日期:2014-04-09 出版日期:2014-12-23 发布日期:2014-12-23
  • 通讯作者: 胡永浩,教授,E-mail:yhh0817@126.com
  • 作者简介:董泓(1987-),男,甘肃临洮人,硕士,主要从事猪瘟病毒蛋白与宿主蛋白的相互作用研究,E-mail:dong.hong55555@163
  • 基金资助:

    国家自然科学基金项目(31201921)

Identification of the Interaction between Classical Swine Fever Virus C Protein and Porcine β-actin

DONG Hong1,LI Dan2,CHEN Jia-ning2,LI Su2,HE Wen-rui2,FENG Shuo2,HE Fan2,LIAO Ya-jin2,SUN Yuan2,HU Yong-hao1*   

  1. (1.College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730000,China;2.State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China)
  • Received:2014-04-09 Online:2014-12-23 Published:2014-12-23

摘要:

为了研究猪瘟病毒(CSFV)和宿主间的相互作用,采用Co-IP方法,利用CSFV C蛋白从PK-15细胞中钓取与其相互作用的宿主蛋白质,经SDS-PAGE电泳,银染,选取差异条带,进行质谱分析,选取β-actin蛋白作为研究的对象。经内源性Co-IP试验和GST pulldown试验进一步证实CSFV C蛋白和β-actin可以特异性结合,激光共聚焦试验表明两者共定位于细胞质。本研究表明CSFV C蛋白与宿主细胞β-actin 蛋白存在相互作用,为深入研究CSFV蛋白质与宿主蛋白质间相互作用提供了实验依据。

Abstract:

To study the interactions between the classical swine fever virus (CSFV) and the host,coimmunoprecipitation (Co-IP) and subsequent mass spectrometry were used to screen and identify the host proteins interacted with CSFV C protein from PK-15 cells.β-actin was identified as a potential interacting partner of C protein.In addition,the interaction of the two proteins was further confirmed by endogenous Co-IP and GST pulldown.Moreover,laser confocal imaging confirmed that β-actin colocalized with the CSFV C protein in the cytoplasm.These results showed that CSFV C interacted with β-actin which would facilitate further study of interactions between CSFV proteins and host cell proteins.

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