羊轮状病毒GS2023株VP7基因的克隆与序列分析  

Cloning and sequence analysis of the VP7 gene of sheep rotavirus GS2023 strain

作　　者：王天宇 赵登率 张远航 李平 于頔浠 赵孟孟 高寒 覃丽梅 郭利伟[1] 张克山 WANG Tianyu;ZHAO Dengshuai;ZHANG Yuanhang;LI Ping;YU Dixi;ZHAO Mengmeng;GAO Han;QIN Limei;GUO Liwei;ZHANG Keshan(College of Animal Science and Technology,Yangtze University,Jingzhou Hubei 434023;College of Animal Science and Technology,Foshan University,Foshan Guangdong 528225)

机构地区：[1]长江大学动物科学技术学院,湖北荆州434023 [2]佛山大学动物科技学院,广东佛山528225

出　　处：《现代牧业》2024年第4期38-47,共10页Modern Animal Husbandry

基　　金：国家重点研发计划项目(2023YFD1801302,2023YFD1801301);佛山大学高层次人才科研启动项目(CGZ07001)。

摘　　要：为研究羊轮状病毒(Rotavirus,RV)VP7基因的结构特点及功能,本试验扩增RV GS2023株VP7基因,并将其连接到pMD18-T载体上,通过生物信息学软件进行序列分析及功能预测。结果表明VP7基因全长978 bp,该蛋白分子质量为37.11763 ku,理论等电点为4.72,偏酸性;不稳定系数为35.29,属于稳定蛋白,不含信号肽。VP7蛋白是一种亲水性蛋白,含有62个糖基化位点和83个磷酸化位点,存在两个跨膜螺旋结构。VP7蛋白最有可能存在于质膜中,高尔基体、过氧物酶体、细胞外基质中也有分布。VP7蛋白的二级结构主要由α螺旋、延伸链、无规则卷曲组成,其中α螺旋占比为33.13%。GS2023株VP7与国内B11-R1株VP7基因相似性最高,为96.6%;GS2023株VP7与同型参考株的VP7氨基酸序列在多个位点有突变,但大部分氨基酸位点是保守的;GS2023株的VP7基因系统发育进化分析结果显示,与同型参考株聚为同一分支,且与B11-R1株亲缘关系最近。In order to understand the sequence characteristics and function of rotavirus VP7 gene,the experiment was conducted to amplify the VP7 gene of RV GS2023 strain based on the existing primers specific for rotavirus VP7 gene and ligated into pMD18-T vector for sequence determination and analysis.The results showed that the amplified VP7 gene was 978 bp in length,the molecular mass of the target protein was 37.11763 ku,the theoretical isoelectric point was 4.72,acidic,and the instability coefficient was 35.29,which was a stable protein without signal peptide.The VP7 protein was a hydrophilic protein,and the VP7 protein contained 62 glycosylation sites and 83 phosphorylation sites,the amino acid distribution of VP7 protein is inside and outside the membrane,with two transmembrane helices.The VP7 protein most likely exists in the plasma membrane,and the rest of the intracellular Golgi apparatus,peroxisomes,and extracellular matrix.the secondary structure of VP7 protein mainly consists ofα-helices,extended strands,and irregular coiled coils,with the proportion ofα-helix accounting for 33.13%.The VP7 of GS2023 strain had the highest homology with the VP7 gene of B11-R1 strain isolated in China,which was 96.6%;the amino acid sequence of the VP7 of GS2023 strain and that of the homozygous reference strain had mutations at several sites,but most of the amino acid sites were conserved;The evolutionary analysis of the VP7 sequence of strain GS2023 showed that it was on the same branch as the homozygous reference strain and had the closest genetic distance to strain B11-R1.

关 键 词：羊轮状病毒 VP7基因 克隆 序列分析 遗传进化分析 

分 类 号：S855.3[农业科学—临床兽医学]