禽呼肠孤病毒C-98、T-98分离株S4基因的克隆与序列分析  

CLONING AND SEQUENCE ANALYSIS OF S4 GENE OF AVIAN REOVIRUS C-98、T-98 ISOLATES

作　　者：党娟[1] 霍金山 何建文[1] 关平原[1] 

机构地区：[1]内蒙古农业大学兽医学院,呼和浩特010018 [2]内蒙古兴安盟动物疫病预防控制中心,乌兰浩特137400

出　　处：《内蒙古农业大学学报（自然科学版）》2012年第2期1-6,共6页Journal of Inner Mongolia Agricultural University(Natural Science Edition)

基　　金：内蒙古自然科学基金资助项目(200308020405)

摘　　要：参考GenBank中禽呼肠孤病毒176株(ARV 176)S4基因序列设计两对引物,分别提取禽呼肠孤病毒T-98和C-98分离株病毒总RNA,应用RT-PCR技术扩增病毒的S4基因,将纯化的目的 DNA与pEASY-T1载体连接、转化后测序。应用计算机软件将所测定序列与参考毒株序列进行比较,结果显示,S4基因核苷酸序列长为1 192bp,均含1个完整的开放性阅读框(24～1 127)。ARV T-98和ARV C-98分离株S4基因的核苷酸同源性很高,为99.9%;与参考毒株ARV 1733、ARV S1133、ARV 750505、ARV 601SI、ARV 919、ARV T6、ARV OS161和DRV YJL S4基因的核苷酸同源性均较高,在99.4%～99.9%之间;与ARV 601G、ARV 1017-1、ARV 916、ARV 918和DRV S14S4基因的核苷酸同源性在80.1%～83.6%之间;与NBV、BRV和MRV 3的S4基因核苷酸同源性在2.4%～53.1%之间。进化树分析显示:11株禽呼肠孤病毒分离株和ARV T-98、C-98分离株的S4基因可分成3个不同的系谱。According to the S4 gene sequence of Avian reovirus strain 176（ARV 176） published in GenBank,two pairs of primers were synthesized.The total RNA of ARV isolate T-98 and C-98 isolated in Tianjin and inner Mongolia were extracted.The S4 gene of virus was amplificated by RT-PCR and then cloned into pESAY-T1 plasmid and then sequenced.The sequences were compared with reference strains gene sequences by computer software.The results showed that S4 genes were 1192bp and contained one ORF（24~1127）.Comparision of nucleotide sequence showed that high homology was 99.9% between ARV T-98 and ARV C-98.The homology of S4 gene was about 99.4%~99.9% compared with reference strains ARV 1733、ARV S1133、ARV 750505、ARV 601SI、ARV 919、ARV T6、ARV OS161 、DRV YJL and 80.1%~83.6% compared with reference strains ARV 601G、ARV 1017-1、ARV 916、ARV 918、DRV S14 and 2.4%~53.1% compared with reference strains NBV、BRV、MRV 3.phylogenetic analysis of S4 gene of 11 ARV strains and ARV T-98、C-98 showed that these genes have separated into three lineages.

关 键 词：禽呼肠孤病毒 S4基因: 克隆 列分析 

分 类 号：S855[农业科学—临床兽医学]