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目的 小反刍兽疫（Peste des petits ruminants, PPR）和山羊痘（Goat pox）是两种危害小反刍动物的高度接触性传染疫病，严重阻碍了我国养羊产业的健康发展。方法 为构建表达小反刍兽疫病毒（Peste des petits ruminants virus, PPRV）HN蛋白的山羊痘重组病毒载体二联疫苗，以先前构建的p UC19T-ΔTK-RFP载体，利用同源重组技术将PPRV HN基因克隆至p UC19T-ΔTK-RFP上构建p UC19T-ΔTK-PPRV-HN转移载体。将转移载体转染Vero细胞，通过与红色荧光山羊痘病毒（GTPV-ΔTK-RFP）重组进行病毒拯救。利用有限稀释法将拯救成功的重组病毒（GTPV-ΔTK-PPRV-HN）进行纯化。再通过蛋白免疫印迹、间接免疫荧光和测序验证GTPV-ΔTK-PPRVHN重组病毒的生物学及其增殖特性。结果 结果表明，本研究成功构建GTPV-ΔTK-PPRV-HN重组病毒并且该重组病毒能够表达PPRV HN蛋白，重组病毒生长特性良好。结论 本研究为PPRV和GTPV的二联疫苗提供理论基础和实践依据，将有利于保障养羊产业的健康发展，助力乡村振兴。
Abstract：Objective Peste des petits ruminants(PPR) and goatpox(GTP) are two highly contagious viral diseases that severely affect small ruminants, posing serious challenges to the sustainable development of the sheep and goat industry in China. Methods To develop a recombinant goatpox virus-vectored bivalent vaccine expressing the hemagglutinin(HN) glycoprotein of PPR virus(PPRV), the previously constructed pUC19T-ΔTK-RFP vector was used as the backbone. The HN gene of PPRV was inserted into pUC19T-ΔTK-RFP via homologous recombination to generate the transfer plasmid pUC19T-ΔTK-PPRV-HN, which was transfected into Vero cells for recombination with a red fluorescent goatpox virus(GTPV-ΔTK-RFP) to rescue the recombinant virus. The rescued recombinant virus(GTPV-ΔTK-PPRV-HN) was purified by limiting dilution, and its expression of PPRV HN was verified by Western blotting, indirect immunofluorescence assay, and sequencing. Results The recombinant GTPV-ΔTK-PPRV-HN was successfully constructed and shown to stably express the PPRV HN protein, exhibiting favorable replication characteristics comparable to the parental strain. Conclusion These findings provide both theoretical and experimental support for the development of a bivalent vaccine against PPRV and GTPV, contributing to improved disease prevention and control in small ruminants and promoting the sustainable development of animal husbandry in support of rural revitalization.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.250257
中图分类号:S852.65
引用信息:
[1]陈韵,任善会,杜新潮,等.表达小反刍兽疫田间毒株HN糖蛋白重组山羊痘毒株的构建及其增殖特性分析[J].病毒学报,2026,42(03):976-986.DOI:10.13242/j.cnki.bingduxuebao.250257.
基金信息:
青海省基础研究计划项目(项目号:2024-ZJ-721),题目:小反刍兽疫病毒糖蛋白(F/H)羊痘活载体基因工程二联疫苗的创制及其应用; 国家自然科学基金地区基金(项目号:32460876),题目:宿主DNA损伤应答反应通过ATR-CHK1信号轴促进羊痘病毒复制的机制研究~~
2025-08-25
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