大熊猫轮状病毒CH-1株VP1基因的克隆和生物信息学分析  

Cloning and bioinformatics analysis of VP1 gene of giant panda rotavirus strain CH-1

作　　者：郭玲[1] 杨绍林[1] 张和民[2] 李德生[2] 王承东[2] 曹三杰[1] 黄晓波[1] 文心田[1] 陈世界 颜其贵[1,4] 

机构地区：[1]四川农业大学动物医学院,四川雅安625014 [2]四川省出入境检疫检验局,四川成都610041 [3]中国保护大熊猫研究中心,四川雅安625000 [4]四川农业大学动物疫病与人类健康四川省重点实验室,四川雅安625014

出　　处：《中国兽医科学》2014年第1期27-32,共6页Chinese Veterinary Science

基　　金：科技部公益性行业科研专项(200910188);四川省科技厅科技支撑计划项目(2010SZ0050);林业公益性行业科研专项(201104050);大熊猫国际合作研究项目(SD0418);四川农业大学优秀硕士论文培育基金资助项目(YSPY1202)

摘　　要：为研究大熊猫源轮状病毒(RV)CH-1株外衣壳蛋白VP1基因的结构及功能,采用MA-104细胞增殖大熊猫源RV,提取总RNA,运用RT-PCR扩增VP1基因,将阳性克隆进行测序,用生物信息学软件预测其功能,并构建系统进化树。结果显示,成功获得了长3 287bp的GPRV VP1蛋白基因(GenBank登录号:HQ641297);经生物信息学分析,此序列包含有一个3 267bp的完整开放阅读框,编码1 089个氨基酸;无信号肽;无跨膜区,其整个蛋白都位于病毒膜外区;抗原表位预测显示VP1蛋白有41个抗原表位;系统进化树分析显示大熊猫轮状病毒CH-1株VP1基因与猪A组轮状病毒VP1基因的进化距离最近。本研究成功获得了大熊猫源RV VP1基因,为今后研究此基因的生物学功能奠定了基础。The VP1 gene of giant panda rotavirus（GPRV） strain CH-1 was amplified by RT-PCR from GPRV-infected MA-104 cells, and sequencing and bioinformatics analysis were then conducted to investi- gate the structure and function of GPRV VP1 protein. The function of this protein was predicted by bio- informatics software and the phylogenetic tree of VP1 gene was constructed. The results indicated that a length of 3 287 bp GPRV VP1 protein encoding gene was obtained （GenBank accession number. HQ641297）. Bioinformatics analysis showed that the sequence contained the complete coding region,which was 3 267 bp,encoding 1 089 aa. The protein had no signal peptide and transmembrane regions and 41 anti- genic determinants were predicted by online analysis. The phylogenetic tree showed that the evolution dis- tance of GPRV VP1 gene was homogeneous to porcine rotavirus A. This research provides some new clues for further studying the biological functions of the gene of the virus.

关 键 词：大熊猫轮状病毒 VP1基因 克隆 生物信息学分析 

分 类 号：S852.659.4[农业科学—基础兽医学]