兔出血症病毒(RHDV)WHNRH株的分离鉴定及基因组全序列的测定与分析  

Isolation,identification and genome sequenceing of a Rabbit hemorrhagic disease virus strain WHNRH

作　　者：李建文[1] 王红宁[1] 田浪[1] 黄勇[1] 张夏兰[1] 

机构地区：[1]四川农业大学动物科技学院动物预防医学生物工程实验室,雅安625014

出　　处：《微生物学报》2006年第5期720-725,共6页Acta Microbiologica Sinica

摘　　要：从发病长毛兔中分离鉴定了兔病毒性出血症病毒WHNRH株。参考GenBank中已登录的RHDV毒株序列对RHDV WHNRH分离株进行了全基因组序列测定与分析。设计5对扩增区段相互重叠的RHDV特异性引物，扩增除5’和3’末端以外的序列，采用设计锚引物的5’RACE方法以及针对RHDV 3’末端的polyA结构设计引物获得了RHDV WHNRH株的5’和3’末端序列。胶回收各PCR产物，连接pMD18-T克隆载体，测得RHDV WHNRH分离株的基因组全长为7437nt（不包括polyA），与GenBank公布的全部共6株RHDV全基因序列进行同源性比较分析，同源性在89．0％～97．1％之间，ORF1同源性为89．0％～97．1％，编码氨基酸序列的同源性为95．2％～98．7％；ORF2的核酸苷序列同源性为92．1％～97．7％，编码氨基酸序列的同源性为94．1％～96．6％。The rabbit hemorrhagic disease virus （RHDV） is a single-stranded positive-sense RNA virus of the family Caliciviridae with a high morbidity and mortality rates of about 90 % in adult rabbits. A strain of rabbit hemorrhagic disease virus named WHNRH was isolated and identified in the research. The genome of WHNRH was then sequenced. Primers were designed referring to the genome sequences of RHDV published in GenBank and a RACE was applied to get the 5＇ terminus sequences. The other terminus sequences of WHNRH was acquired referring to the genome＇s polyA structure of RHDV.The genome was amplified with RT-PCR. All the RT-PCR products were cloned into the pMD18-T vector and sequenced. The sequencing result indicated that the complete genome of RHDV isolated strain WHNRH was composed of 7437nt （not including polyA of 3＇ terminus）, the identities were between 89.4% and 97.1% compared with the published genome sequences of six RHDV strains. The ORF1 of the genome of WHNRH was between 10nt and 7044nt and a polypeptide with 2344 amino acids was coded, the identities of nucleotide and amino acids sequences were 89.1% - 96.1% and 96.0% - 98.4% respectively compared with the six published RHDV strains. The ORF2 of the genome of WHNRH was between 7025nt and 7378nt and a polypeptide with 177 amino acids was coded, the identities of nucleotide and amino acids sequences were all between 92.1% and 96.9 % compared with the six published RHDV strains.

关 键 词：分离鉴定 兔出血症病毒 全基因组测序 WHNRH 

分 类 号：S852.65[农业科学—基础兽医学]