Screening of host proteins interacting with GP, VP30, NP,and L proteins of Ebola virus by yeast two-hybrid system
Author(s): LI Cheng-gong, ZHANG Bu-chang, GAO Ting, CAO Cheng
Pages: 249-255
Year: 2017 Issue: 4
Journal: Bulletin of the Academy of Military Medical Sciences
Keyword: Ebola virus; yeast two-hybrid technique;
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.
Pages: 249-255
Year: 2017 Issue: 4
Journal: Bulletin of the Academy of Military Medical Sciences
Keyword: Ebola virus; yeast two-hybrid technique;
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.
Related Articles