[Objective] The paper was to obtain host proteins interacting with porcine epidemic diarrhea virus (PEDV) N protein. [Method] The re-combinant vector pGBKT7-N of PEDV N gene was constructed and used as the bait plas...[Objective] The paper was to obtain host proteins interacting with porcine epidemic diarrhea virus (PEDV) N protein. [Method] The re-combinant vector pGBKT7-N of PEDV N gene was constructed and used as the bait plasmid to screen the proteins interacting with N protein ofPEDV from the cDNA library of porcine alveolar macrophage (PAM) by yeast two-hybrid method. [Result] There was no toxicity and self activationof bait protein in yeast hybridization system, and six proteins (FTH1, LGALS3, CORO1C, SNRPG, KRTAP5-3, ZNF598) interacting with N proteinwere indentified. It was confirmed that LGALS3 and SNRPG had specific interaction with N protein by return experiment and co-immunoprecipitation(CoIP) test. [Conclusion] The study lays a foundation for further studying the function of PEDV N protein and the pathogenic mechanism of PEDV.展开更多
基金Supported by National Natural Science Foundation of China(31201915,31502071)Key Project of Science and Technology Promoting Agriculture in Shanghai City[HNKGZ(2013)No.3-6,No.5-5]
文摘[Objective] The paper was to obtain host proteins interacting with porcine epidemic diarrhea virus (PEDV) N protein. [Method] The re-combinant vector pGBKT7-N of PEDV N gene was constructed and used as the bait plasmid to screen the proteins interacting with N protein ofPEDV from the cDNA library of porcine alveolar macrophage (PAM) by yeast two-hybrid method. [Result] There was no toxicity and self activationof bait protein in yeast hybridization system, and six proteins (FTH1, LGALS3, CORO1C, SNRPG, KRTAP5-3, ZNF598) interacting with N proteinwere indentified. It was confirmed that LGALS3 and SNRPG had specific interaction with N protein by return experiment and co-immunoprecipitation(CoIP) test. [Conclusion] The study lays a foundation for further studying the function of PEDV N protein and the pathogenic mechanism of PEDV.