摘要
为研究马传染性贫血病毒(EIAV)感染其主要靶细胞马巨噬细胞(eMDM)后与细胞蛋白的相互作用,本研究采用EIAV强毒株EIAVDLV34感染48 h后的马外周血单核细胞分化的eMDM,并以未感染eMDM为对照,提取细胞的蛋白质样品,进行双向凝胶电泳(2-DE)分离,并分析凝胶中差异蛋白点。结果共检测出19个表达差异的蛋白点(ratio>1.4,p<0.05),其中感染组相对于对照组有7个蛋白质上调表达,12个蛋白质呈现下调表达。将差异蛋白进行串联质谱分析进行鉴定,并通过生物信息学方法对这19个差异表达蛋白进行了蛋白互作用分析。此外,对其中5个比较重要蛋白的mRNA水平进行了荧光定量PCR分析,其表达变化与2-DE结果一致。本研究为进一步分析EIAV与其宿主细胞eMDM的相互作用提供了参考。
To investigate the interaction between equine infectious anemia virus （EIAV） and the host equine monocyte-derived macrophages （eMDM）, the differentially expressed proteins in eMDM infected with an EIAV pathogenic strain EIAVDLV34 were analyzed by proteome. Total proteins were extracted from cells infected for 48 hour and the differentially expressed proteins were screened by using 2-D acrylamide gel electrophoresis. Cell extracts from mock-infected eMDM were used as the control. Total 19 differentially expressed proteins （ratio〉l.4, p〈0.05） in EIAV-infected eMDM were found, among which 7 were up-regulated and 12 were down-regulated. All these 19 differentially expressed proteins were identified by examining with tandem mass spectrum. In addition, these proteins were analyzed for possible interactions using several bioinformatics methods. The mRNA levels of five functionally important differentially expressed proteins were further confirmed by quantitative real-time PCR. This study provided important information for further investigation of the interaction between EIAV and its target cells.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2012年第12期933-936,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
中央级公益性科研院所基本科研业务费专项(0302012012)
国家十二五科技重大专项(2012ZX10001-008)
关键词
马传染性贫血病毒
eMDM
差异蛋白
双向电泳
生物信息学
equine infectious anemia virus
eMDM
differentially expressed proteins
two dimensional gel electrophoresis
bioinformatics
