摘要
探究非洲猪瘟病毒(ASFV)RNA解旋酶A859L的基本生物学特征,为后续研究A859L的生物学功能以及A859L在ASFV复制和致病机制中的作用奠定基础。首先,在NCBI Virus数据库中下载不同ASFV毒株的A859L基因序列,构建系统进化树。其次,利用在线软件分析A859L的理化性质和结构。最后,通过构建A859L的真核表达质粒,利用Western-blot和间接免疫荧光试验检测A859L的细胞定位和结合dsRNA的能力。系统进化树显示,A859L在不同ASFV毒株基因组中是一个保守基因,且A859L是一个亲水且不含有跨膜结构域的病毒蛋白。A859L蛋白二级结构中含有α-螺旋(49.65%)、无规则卷曲(37.88%)和延伸链(10.14%)。Western-blot和间接免疫荧光试验结果显示,A859L蛋白主要定位于细胞质且具有结合dsRNA的功能。本研究证实A859L是一个保守基因,揭示A859L蛋白定位于细胞质且具有结合dsRNA的能力,为后续探究A859L蛋白在基因组转录和复制中的作用奠定了基础。
The aim is to investigate the basic biological characteristics of the African swine fever virus(ASFV)RNA helicase A859L,laying the foundation for subsequent studies of its biological functions and its role in ASFV replication and pathogenesis.First,the A859L gene sequences of different ASFV strains were downloaded from the NCBI Virus database,and a phylogenetic tree was constructed.Second,the physicochemical properties and structure of A859L were analyzed using online software.Finally,an eukaryotic expression plasmid for A859L was constructed,and the cellular localization and dsRNA-binding ability of A859L were detected through Western-blot and indirect immunofluorescence assay.The phylogenetic tree revealed that A859L was a conserved gene in the genomes of various ASFV strains,and A859L was a hydrophilic viral protein without a transmembrane domain.The secondary structure of the A859L protein containsα-helices(49.65%),random coils(37.88%),and extended chains(10.14%).Western-blot and indirect immunofluorescence assay results showed that the A859L protein was primarily localized in the cytoplasm and had the ability to bind dsRNA.In conclusion:this study confirms that A859L is a conserved gene,reveals the cytoplasmic localization of the A859L protein and its dsRNA-binding capability,and provides a foundation for further exploring the role of the A859L protein in genome transcription and replication.
作者
周万卉
武正前
张旭辉
周思妮
杨文萍
郑海学
王雅莉
朱向涛
ZHOU Wanhui;WU Zhengqian;ZHANG Xuhui;ZHOU Sini;YANG Wenping;ZHENG Haixue;WANG Yali;ZHU Xiangtao(School of Biological and Pharmaceutical Engineering,Lanzhou Jiaotong University,Lanzhou 730070,China;Key Laboratory of Microbial Resources Exploitation and Application of Gansu Province,Lanzhou 730070,China;State Key Laboratory of Animal Disease Control and Prevention/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;Gulang County Agricultural Industrialization Service Center,Wuwei 733000,China)
出处
《中国兽医科学》
北大核心
2026年第2期193-199,共7页
Chinese Veterinary Science
基金
甘肃省高校教师创新基金项目(2025B-064)
动物疫病防控全国重点实验室开放基金项目(SKLADCPKFKT202408)。
