中国兽医科学2026，Vol.56Issue(5)：595-601,7.DOI:10.16656/j.issn.1673-4696.2026.0102
共表达鸡传染性贫血病毒VP1、VP2基因和鸡γ干扰素基因的重组鸡痘病毒的构建与鉴定
Construction and identification of a recombinant fowlpox virus co-expressing VP1 and VP2 genes of chicken infectious anemia virus and chicken interferon-gamma gene
摘要
Abstract
To construct a recombinant fowlpox virus transfer vector co-expressing the VP1 and VP2 genes of chicken infectious anemia virus(CIAV)and chicken interferon-gamma(ChIFN-γ),a homologous recombination strategy was employed.The VP1 and VP2 genes of CIAV,the ChIFN-γ gene,and the reporter gene lacZ were integrated into a non-essential region for viral replication within the genome of the fowlpox virus(FPV)S-FPV-017 strain.The VP1,VP2,and ChIFN-γ gene expression cassettes controlled by FPV early/late promoter LP2EP2 and lacZ reporter gene expression cassette controlled by P11 promoter(late promoter)were inserted into the non-essential region of FPV genome(S-FPV-017).The recombinant transfer plasmid pSY681-VP1-VP2-ChIFN-γ was transfected into chicken embryo fibroblasts(CEF)pre-infected with parental FPV strain S-FPV-017 for homologous recombination.After 8 rounds of plaque screening and purification,the recombinant virus rFPV-VP1-VP2-ChIFN-γ expressing CIAV VP1,VP2,ChIFN-γ and lacZ reporter gene was successfully obtained.PCR identification confirmed that CIAV VP1,VP2,and ChIFN-γ genes were integrated into the recombinant virus genome.Indirect immunofluorescence assay and Western-blot results demonstrated that the corresponding proteins were effectively ex-pressed in CEF cells infected with the recombinant virus.Compared with the parental virus,the recombi-nant virus showed no significant differences in replication characteristics and cytopathic effects on CEF cells.These findings demonstrate that multiple exogenous genes from avian pathogens can be simul-taneously inserted and expressed in a non-essential region of FPV genome,providing important founda-tion for development of multivalent recombinant viral vector vaccines.关键词
鸡痘病毒/鸡传染性贫血病毒/VP1基因/VP2基因/ChIFN-γ基因Key words
fowlpox virus/chicken infectious anemia virus/VP1 gene/VP2 gene/ChIFN-γ gene分类
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周小汇,刘丹,薛麒,孔冬妮,吕岱玥,印春生,毛娅卿,湛洋..共表达鸡传染性贫血病毒VP1、VP2基因和鸡γ干扰素基因的重组鸡痘病毒的构建与鉴定[J].中国兽医科学,2026,56(5):595-601,7.基金项目
国家自然科学基金项目(32573356,32202790) （32573356,32202790）
"十四五"国家重点研发计划项目(2023YFD1800603) （2023YFD1800603）
2022年度湖南省"三尖"创新人才工程项目(2022RC1046) （2022RC1046）
