羊轮状病毒GS2023株NSP2基因克隆及序列分析  

Cloning and sequence analysis of NSP2 gene of sheep rotavirus GS2023 strain

作　　者：于頔浠 赵登率 张远航 李平 王天宇 张克山 高寒 覃丽梅 赵孟孟 李家奎 YU Dixi;ZHAO Dengshuai;ZHANG Yuanhang;LI Ping;WANG Tianyu;ZHANG Keshan;GAO Han;QIN Limei;ZHAO Mengmeng;LI Jiakui(Tibetan Academy of Agriculture and Animal Sciences,Nyingchi 860114,China;College of Animal Science and Technology,Foshan University,Foshan 528225,China)

机构地区：[1]西藏农牧学院动物科学学院,西藏林芝860114 [2]佛山大学动物科技学院,广东佛山528225

出　　处：《畜牧与兽医》2025年第4期121-129,共9页Animal Husbandry & Veterinary Medicine

基　　金：国家重点研发计划项目(2023YFD1801302,2023YFD1801301);国家肉牛牦牛产业技术体系项目(CARS-37);佛山大学高层次人才研究创业项目(CGZ07001)。

摘　　要：为研究羊轮状病毒NSP2蛋白的结构和功能,以羊轮状病毒GS2023株cDNA为模板对NSP2基因进行PCR扩增与克隆,并利用多种生物信息学分析预测软件分析NSP2的生物信息学特征。结果:NSP2基因的开放阅读框长度为954 bp,编码317个氨基酸;NSP2属于亲水性、稳定蛋白,不存在信号肽;亚细胞定位预测表明其主要定位于细胞质和细胞核中,属于非跨膜蛋白;该蛋白含有36个糖基化位点以及49个磷酸化位点;二级结构由α螺旋结构、延伸链和无规则卷曲构成,三级结构则呈自身首尾相连结构;NSP2蛋白与国内外轮状病毒株的氨基酸相似性为90.2%~98.7%;GS2023株与国内牛源轮状病毒株SCMY1和SCMY2氨基酸对比发现在第38位由赖氨酸突变为精氨酸,与国内多种羊源毒株和牛源毒株氨基酸对比发现在第100、202位分别由丝氨酸突变为天冬酰胺、由缬氨酸突变为异亮氨酸;系统发育进化树分析结果显示,GS2023株与国内牛源轮状病毒株遗传距离最近,与其他羊源轮状病毒遗传距离较远。本研究成功克隆羊轮状病毒NSP2基因并分析NSP2蛋白的生物学特征,为进一步研究NSP2蛋白的结构和功能提供理论参考。In order to study the structure and function of sheep rotavirus NSP2 protein,the cDNA of a sheep rotavirus GS2023 strain was used as a template to amplify and clone the NSP2 gene by PCR,and various bioinformatics analyses and prediction software were used to ana⁃lyze the bioinformatics characteristics of NSP2.The results showed that the open reading frame of the NSP2 gene was 954 bp,encoding 317 amino acids.NSP2 was a hydrophilic and stable protein,with no signal peptide.The prediction of subcellular localization showed that it was mainly located in cytoplasm and nucleus and belonged to non-transmembrane protein.The NSP2 protein contained 36 glycosylation sites and 49 phosphorylation sites.Its secondary structure was composed ofαhelix structure,extended chain and random curl,while its tertiary struc⁃ture was connected end to end.The amino acid similarity between the NSP2 protein and the rotavirus strains in China and in the world was 90.2%-98.7%.Compared with bovine rotavirus strains SCMY1 and SCMY2 in China,the amino acids of GS2023 were mutated from lysine to arginine at position 38,and from serine to asparagine and from valine to isoleucine at positions 100 and 202,respectively.The phylogenet⁃ic tree analysis showed that the genetic distance between the GS2023 strain and the bovine rotavirus strains in China was the closest,but it was far from other sheep rotaviruses.In this study,the NSP2 gene of a sheep rotavirus was successfully cloned and the biological characteris⁃tics of the NSP2 protein were analyzed,which provided theoretical reference for further research on the structure and function of the NSP2 protein.

关 键 词：羊 轮状病毒 NSP2 序列分析 系统发育进化分析 

分 类 号：S852[农业科学—基础兽医学]