免疫共沉淀联合质谱技术筛选宿主因子CypB的互作蛋白

发布时间：2018-03-17 00:25

本文选题：羊传染性脓疱病毒　切入点：CypB　出处：《中国兽医学报》2017年05期 　论文类型：期刊论文

【摘要】：为探讨亲环蛋白B(cyclophilin B,CypB)促进羊传染性脓疱病毒(orf virus,ORFV)复制增殖的机制,以ORFV早期感染MDBK细胞为试验材料,利用免疫共沉淀技术筛选与CypB互作的蛋白。免疫共沉淀产物的Western blot和SDS-PAGE的分析结果显示,在大小约为25 000处可见有明显的差异条带。切下该差异蛋白条带进行质谱鉴定分析,将获得的原始质谱数据使用Mascot软件中的串级质谱数据搜索功能(MS/MS Ions Search)进行搜索鉴定,初步筛选获得6个可能与CypB相互作用的病毒相关蛋白。为进一步深入研究CypB在ORFV早期侵染过程中的作用奠定基础。
[Abstract]:In order to investigate the mechanism of cyclophilin BypB) in promoting the replication and proliferation of sheep infectious pustules virus (ORFV), the early infection of MDBK cells with ORFV was used as the experimental material. The immunoprecipitation technique was used to screen proteins interacting with CypB. The results of Western blot and SDS-PAGE analysis of the immunoprecipitation products showed that there were obvious differential bands at about 25 000. The differential protein bands were cut off and identified by mass spectrometry. The original mass data will be searched and identified using the Cascade MS Ions search function in the Mascot software. Six virion related proteins which may interact with CypB were obtained, which laid a foundation for further study on the role of CypB in the early infection of ORFV.
【作者单位】：吉林大学动物医学学院;
【基金】：国家自然科学基金资助项目(31672554) 吉林省重点科技攻关项目(20140204076NY) 吉林省世行基金资助项目(2011-Y20)
【分类号】：S852.65

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