中国动物传染病学报2026，Vol.34Issue(3)：26-33,8.
表达非洲猪瘟病毒pB602L的JEV复制子构建及鉴定
Construction and Characterization of a Japanese Encephalitis Virus Replicon Expressing ASFV pB602L
摘要
Abstract
Given that RNA virus replicons can achieve high levels of expression of exogenous antigenic genes,the African swine fever virus(ASFV)B602L gene was selected for attempt to construct a Japanese encephalitis virus(JEV)replicon expressing ASFV pB602L in the present study.The replicon expression was verified by transient transfection and construction of replication-defective virus.Firstly,codon optimization of the ASFV B602L gene was performed and the optimized B602L gene was cloned into the JEV replicon(pJEV-CMV-EGFP)to obtain the replicon pJEV-CMV-pB602L.Subsequently,pJEV-CMV-pB602L was transfected into HEK-293T cells and expression of pB602L and JEV non-structural proteins were analyzed by indirect immunofluorescence and Western blot.The results showed that pJEV-CMV-pB602L was able to express the B602L gene in the cells while the non-structural protein NS1 in the JEV replicon backbone was also expressed.To further use this replicon to construct a replication-defective JEV expressing pB602L,pJEV-CMV-pB602L was co-transfected into BHK-21 cells with the plasmid pCAGGS-CprME for expression of the structural proteins of JEV and construction of replication-defective JEV expressing pB602L(rJEVΔC/prM/E-pB602L).The results showed that the defective virus expressing pB602L was successfully constructed as demonstrated in indirect immunofluorescence and Western blot.In conclusion,a JEV replicon expressing ASFV pB602L was successfully constructed in this study and both transient transfection and recombinant defective virus infection showed that this replicon effectively expressed the ASFV pB602L.These results laid the foundation for the subsequent construction of replication-defective JEV expressing other ASFV antigens.关键词
JEV复制子/非洲猪瘟病毒/B602L/复制缺陷病毒Key words
JEV replicon/African swine fever virus/B602L/replication-defective virus分类
农业科技引用本文复制引用
程星雨,邱亚峰,肖长广,周群,李宗杰,邵东华,刘珂,李蓓蓓,魏建超,马志永..表达非洲猪瘟病毒pB602L的JEV复制子构建及鉴定[J].中国动物传染病学报,2026,34(3):26-33,8.基金项目
国家重点研发计划子课题(2021YFD1801401-1) （2021YFD1801401-1）
