重庆两株H5N1流感病毒的HA、NA基因克隆及序列分析  

HA and NA Gene Clone and Sequence Analysis of Two H5N1 Virus Isolates from Chongqing

作　　者：胡仁建[1,2] 顾盼[1] 蔡家利[1] 刘海军[3] 丁森[4] 张奎[2] 李重阳[2] 陈华 王冬梅[6] HU Ren-jian GU Pan CAI Jia-li LIU Hai-jun DING Sen ZHANG Kui LI Chong-yang CHEN Hua WANG Dong-mei(College of Pharmacy and Biological Engineering, Chongqing University of Technology, Chongqing 400074, China State Key Laboratory of Silkworm Genome Biology, Southwestern University, Chongqing 400074, China Chongqing City Public Security Bureau, Chongqing 400082, China Nanjing University, Nanjing 210023, China Zhuhai United Laboratories Co. Ltd. , Guangzhou 528400, China China Agricultural University, Beijing 100193, China)

机构地区：[1]重庆理工大学药学与生物工程学院,重庆400074 [2]西南大学家蚕基因组生物学国家重点实验室,重庆400074 [3]重庆市公安局,重庆400074 [4]南京大学,南京210023 [5]珠海联邦制药有限公司,广州528400 [6]中国农业大学,北京100193

出　　处：《重庆理工大学学报（自然科学）》2017年第1期67-74,共8页Journal of Chongqing University of Technology：Natural Science

基　　金：重庆市科技攻关计划项目(cstc2012gg-yyjs80014);重庆市科技人才培养计划项目(cstc2013kjrc-1jrcpy80001)

摘　　要：2012年,从重庆北碚区和巴南区的家禽养殖场死鸭和病鸡体内分离出两株H5N1禽流感病毒,分别命名为A/duck/Chongqing/BB/H5N1和A/chicken/Chongqing/BN/H5N1,并对这两株H5N1禽流感病毒的HA、NA和M蛋白的基因进行了克隆及序列分析,在基因和蛋白质水平上分析其同源性。遗传进化树分析结果表明:这两毒株的HA、NA和M基因不处于同一支上,亲缘关系较远;这两毒株的HA、NA、M蛋白的基因同源性分别达95%,95%,97%;两毒株的HA蛋白裂解位点氨基酸序列均为RERRR-KR,符合高致病性禽流感的特征。本次分离的流感病毒的基因克隆鉴定和序列分析为禽流感的流行病学研究、疫苗研制以及防控提供重要理论依据。Avian influenza（AI）,a virus subtype H5N1 virus is a highly pathogenic avian influenza（HPAI）,which is panzootic in poultry,then continues to spread and pose a major challenge to animal and human health.In 2012,two outbreaks of HPAI virus occurred and we isolated two H5N1 virus strains from two poultry farms in the Chongqing Beibei District and Banan District,named A/duck/Chongqing/BB/2012 and A/chicken/Chongqing/BN/2012,respectively.In this study,the hemagglutinin（HA）,neuraminidase（NA） and matrix protein（M） genes of these two strains were cloned and sequenced,and then the homogeneity and the heterogeneity were analyzed at the genetic and proteinic levels.The results shows that the homogeneity of the HA,NA and M genes was 95%,95% and 97%,respectively.The phylogenetic analyses of HA,NA and M genes showed that A/duck/Chongqing/BB/2012 and A/chicken/Chongqing/BN/2012 were distant relatives.However,both of the two strains have the same HA gene proteolytic cleavage sites,RERRR-KR,which accorded with the molecular characteristic of high pathogenic AIV.These results could provide more information for the analysis of the sequence of avian influenza virus H5N1,including variation analysis,evolutionary analysis,as well as the development of vaccine.

关 键 词：禽流感病毒 H5N1 基因克隆 序列分析 进化分析 

分 类 号：S855.3[农业科学—临床兽医学]