野鸭源H6N1亚型禽流感病毒全基因的克隆及序列分析  

Cloning and sequencing analysis of H6N1 subtype AIV strains isolated from mallard

作　　者：丁洁[1,2] 桑晓宇[2] 程凯慧[2] 于志君[2] 张坤[2] 柴洪亮[1] 王铁成[2] 高玉伟[2] 夏咸柱[2] 华育平[1] 

机构地区：[1]东北林业大学,黑龙江哈尔滨150010 [2]军事医学科学院军事兽医研究所,吉林长春130062

出　　处：《中国兽医学报》2013年第3期394-399,共6页Chinese Journal of Veterinary Science

基　　金：国家科技支撑计划资助项目(2010BAD04B01);国家林业局野生动物疫源疫病监测资助项目(2010)

摘　　要：扩增、克隆H6N1亚型禽流感病毒(AIV)A/mallard/Sanjiang/275/2007(M-275)的全基因序列,进行序列分析,并探讨该病毒的进化特点。设计特异性引物,扩增克隆2007年从三江自然保护区野鸭身上分离到的H6N1亚型AIV,测序后进行序列分析。同源性分析发现,M-275与H6亚型和N1亚型AIV同源性最高,说明M-275为H6N1亚型。其裂解位点为PQIETR↓G,无连续碱性氨基酸的插入,说明M-275属于典型的低致病性毒株。通过序列比对,M-275的HA基因与2000年以后北美以及中国地区分离到的H6亚型AIV同源性在97%以上,与HA基因同源性最高的毒株为A/northern pintail/Alaska/44202-143/2006(H6N1)和A/northern pintail/Alaska/44204-158/2006(H6N4),与NA基因的核苷酸同源性最高的毒株是A/mallard/ON/499/2005(H5N1),推测M-275的NA基因片段来自于该病毒谱系。进化分析结果显示,HA基因与美国、日本、中国等国家和地区流行的N1、H2、N4、N8和N9等亚型毒株处于同一分支;NA基因与美国、日本等国家和地区流行的H2、H5、H6、H11、H3等亚型毒株处于同一分支;PB1基因与美国、日本等国家和地区流行的毒株处于同一分支;而PB2、NP、M以及NS基因则与日本、韩国、香港和中国地区的病毒株处于同一分支;PA同中国地区和澳大利亚、马来西亚、瑞典等国家的毒株处于同一分支。M-275毒株基因组构成来源复杂,NA基因和NP基因分别与野鸭源的H5N1和H1N1等毒株同源性很高,表明H6亚型禽流感病毒也在不断的发生重排,值得深入研究。To study the evolution characteristic of M-275 strain ,we amplified,cloned and analyzed the full-length sequence of H6N1 eight segments. Design specific primers and cloned the eight segments of H6N1 AIV which was isolated from mallard in Sanjiang, then sequencing analysis. By homology analysis,M-275 shared the highest homology with H6 subtype and N1 subtype,so M-275 is H6N1 subtype. The cleavage site is PQIETR ↓ G, without continuous alkaline amino acid insertion,so M-275 is a typical low pathogenic strain. By sequence alignment,the HA gene of M- 275 has above 97% homology with the H6 subtype virus isolated in North America and China af- ter 2000, HA gene of M-275 shared the highest homology with A/Northern pintail/Alaska/44202- 143/2006 （H6N1） and A/Northern pintail/Alaska/44204-158/2006 （H6N4）, and NA gene shared the highest homology with A/mallard/ON/499/2005 （H5N1）. The phylogenetic analysis showed that HA gene of M-275 originated from the N1, H2, N4, N8 and N9 subtype viruses in America and Japan,NA gene from the H2, HS, H6, Hll, H3 subtype viruses in America and Japan, PB1 gene from the viruses in America and Japan, and PB2, NP, M and NS genes from Japan, South Ko- rea, Hong kong and China,PA from the viruses in China, Malaysia, Sweden and Australia. Thisstrain has a diversity source, HA gene and NA gene have highly homology with the mallard source H5N1 and H1N1 subtype , showed that H6 subtype influenza virus were constantly undergoing rearrangement.

关 键 词：禽流感病毒 H6N1 序列分析 

分 类 号：S852.65[农业科学—基础兽医学]