黑龙江八一农垦大学学报2025，Vol.37Issue(2)：46-52,7.DOI:10.3969/j.issn.1002-2090.2025.02.007
PEDV S1-NTD和S1-CTD不同组合假病毒的构建及鉴定
Construction and Characterization of Pseudoviruses with Different Combinations of PEDV S1-NTD and S1-CTD
摘要
Abstract
Porcine epidemic diarrhea virus(porcine epidemic diarrhea virus,PEDV)is a highly contagious coronavirus that causes severe diarrhea and high mortality in piglets.The N-terminal structural domain(S1 subunit N-terminal domain,S1-NTD)and the C-terminal structural domain(S1 subunit C-terminal domain,S1-CTD)of the S1 subunit of the PEDV spike(spike,S)protein play important roles in viral invasion,but the mechanisms have not been fully elucidated.In order to further explore the synergistic effect of PEDV S1-NTD and S1-CTD in the process of virus invasion into host cells.In this study,S1-NTD and S1-CTD pseudoviruses with different combinations of S proteins of the PEDV classical strain CV777 and the PEDV type GIIa variant strain HM2017 were constructed by using the vesicular stomatitis virus(vesicular stomatitis virus,VSV)pseudovirus system.They were named rvCV777-S1-NTD+CV777-S1-CTD,rvCV777-S1-NTD+HM2017-S1-CTD,rvHM2017-S1-NTD+CV777-S1-CTD,and rvHM2017-S1-NTD+HM2017-S1-NTD.The constructed recombinant virus was identified.The results showed that the different combinations of S protein pseudoviruses observed by transmission electron microscopy(TEM)all presented typical bullet-shaped virus particles with a diameter of approximately 180 nm.Western blot results showed that pseudoviruses expressing different S1-NTD and S1-CTD combinations of S proteins reacted specifically with monoclonal antibodies to the PEDV S protein,with a band size of 180 kDa.Indirect immunofluorescence assay showed that the experimental group all showed clear green fluorescence.The above results proved that the S protein pseudovirus of different S1-NTD and S1-CTD combinations was successfully constructed in this study,providing a experimental material basis for the study of PEDV invasion mechanism.关键词
猪流行性腹泻病毒/水疱性口炎病毒/S1-NTD/S1-CTD/假病毒Key words
porcine epidemic diarrhea virus/vesicular stomatitis virus/S1-NTD/S1-CTD/pseudovirus分类
畜牧业引用本文复制引用
张永晨,齐珊珊,李春秋,朱庆贺,邢晓旭,孙东波..PEDV S1-NTD和S1-CTD不同组合假病毒的构建及鉴定[J].黑龙江八一农垦大学学报,2025,37(2):46-52,7.基金项目
国家自然科学基金区域创新发展联合基金项目(U23A20236),黑龙江省重点研发计划指导项目(GZ20220029),黑龙江省自然科学基金优秀青年项目(YQ2022C028),黑龙江八一农垦大学研究生创新科研项目(YJSCX2022-Y37). （U23A20236）
