军事医学2017，Vol.41Issue(4)：249-255,7.DOI:10.7644/j.issn.1674-9960.2017.04.001
运用酵母双杂交系统筛选与埃博拉病毒GP、VP30、NP、L蛋白存在相互作用的宿主蛋白
Screening of host proteins interacting with GP, VP30, NP,and L proteins of Ebola virus by yeast two-hybrid system
摘要
Abstract
Objective To study the biological function of GP,VP30,NP and L proteins in Ebola virus(EBOV) infectious diseases,and to screen the host proteins which interact with GP,VP30,NP,and L proteins of EBOV from human liver cDNA library using yeast two-hybrid technique.Methods Recombinant PCR was used to construct bait plasmids pGBKT7-GP,pGBKT7-VP30,pGBKT7-NP and pGBKT7-L.Bait strains were combined with human liver cDNA library strains.The positive clones were analyzed by DNA sequencing and bioinformatics.A yeast recovery experiment was performed to further verify and exclude false positive results.Results We constructed bait plasmids pGBKT7-GP,pGBKT7-VP30,pGBKT7-NP and pGBKT7-L with the recombinant PCR method.Six host proteins which could interact with GP,VP30,NP,and L proteins were screened,including COMMD1,ALB,PSMD8,APOA2,CYP2E1,and HP.The yeast recovery experiment proved that COMMD1 and APOA2 might interact with NP protein.Conclusion A number of prey proteins which interact with GP,VP30,NP,and L proteins of EBOV are screened,which may provide reference for the research of EBOV.关键词
埃博拉病毒/酵母双杂交技术/COMMD1/APOA2Key words
Ebola virus/yeast two-hybrid technique/COMMD1/APOA2分类
医药卫生引用本文复制引用
李成功,张部昌,高婷,曹诚..运用酵母双杂交系统筛选与埃博拉病毒GP、VP30、NP、L蛋白存在相互作用的宿主蛋白[J].军事医学,2017,41(4):249-255,7.基金项目
国家自然科学基金应急管理资助项目(81550001) （81550001）
国家自然科学基金重大资助项目(81590766) （81590766）
病原微生物生物安全国家重点实验室开放研究基金资助项目(SKLPBS1507) （SKLPBS1507）
