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目的 探索并开发新一代高效轮状病毒疫苗，为低收入国家易感人群的轮状病毒防治提供实验依据。方法 基于G6P[1]型牛轮状病毒BLL株体外高效复制特性及全基因组测序结果，利用同源重组等技术构建了包含BLL株11个基因片段的感染性克隆质粒，并通过反向遗传学技术拯救该病毒株。采用dsRNA-PAGE硝酸银染色和RT-PCR扩增技术鉴定毒株，并进一步通过间接免疫荧光、qRT-PCR技术测定拯救毒株的病毒滴度及生长动力学。结果 结果表明构建的G6P[1]型牛轮状病毒BLL株感染性克隆可以成功拯救，拯救病毒株的病毒滴度及复制能力均高于羊轮状病毒（Lanzhou lamb rotavirus, LLR）疫苗株。结论 本研究成功建立了G6P[1]型牛轮状病毒BLL株的感染性克隆及病毒拯救体系，为进一步开展基于牛轮状病毒BLL株的疫苗研发及相关机制研究提供了技术手段和基础。
Abstract：Objective To explore and develop a new generation of highly effective rotavirus vaccines and to provide an experimental basis for the prevention and control of rotavirus infection among susceptible populations in low-income countries. Methods Based on the high replication efficiency in vitro and whole-genome sequencing results of the G6P[1] bovine rotavirus BLL strain, infectious clone plasmids containing all 11 genomic segments of the BLL strain were constructed using homologous recombination and related techniques, and the virus was rescued by reverse genetics. The rescued virus was identified by dsRNA-PAGE silver staining and RT-PCR amplification. In addition, indirect immunofluorescence and qRT-PCR were used to determine viral titer and growth kinetics of the rescued virus. Results The results showed that the infectious clone of the G6P[1] bovine rotavirus BLL strain could be successfully rescued, and that the rescued virus exhibited higher viral titer and replication capacity than the Lanzhou lamb rotavirus(LLR) vaccine strain. Conclusion In this study, an infectious clone and virus rescue system for the G6P[1] bovine rotavirus BLL strain was successfully established, providing technical support and a foundation for further vaccine development and related mechanistic studies based on the bovine rotavirus BLL strain.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.250200
中图分类号:R373.2
引用信息:
[1]魏潇,余俊杰,李轲,等.G6P[1]型牛轮状病毒BLL株感染性克隆的构建[J].病毒学报,2026,42(02):481-492.DOI:10.13242/j.cnki.bingduxuebao.250200.
基金信息:
国家自然科学基金（项目号：21934005）,题目：中国母乳糖组分离分析及与轮状病毒蛋白相互作用研究~~
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