新疆羊口疮病毒分离鉴定及B2L基因分析与表达  

Isolation of Orf virus in Xinjiang and prokaryotic expression of the B2L gene

作　　者：李瑞芳[1] 李国华[1] 孟仁[1] 乔军[1] 张辉[1] 陈创夫[1] 

机构地区：[1]石河子大学动物科技学院,新疆石河子832000

出　　处：《中国预防兽医学报》2013年第3期202-205,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家973计划(2010CB30203);国际科技合作项目(2006DFA33740)

摘　　要：为研究新疆地区羊口疮(Orf)病毒(ORFV)的生物学特性及流行特征,本研究采集新疆地区疑似Orf的羔羊结痂病料,用MDBK传代细胞进行病毒分离培养及电镜观察,进行动物回归实验;对ORFV B2L基因进行PCR扩增,并构建B2L基因原核表达重组质粒pET-32a-B2L,转化至大肠杆菌BL21。将表达产物进行SDS-PAGE及western blot检测,结果证明所获得的分离株ORFV-shz为ORFV,与India 67/04分离株的亲缘关系最近,其重组B2L蛋白大小为60 ku,并具有良好反应原性。To investigate the biological and epidemiological characters of Orf virus （ORFV）, the dry scab samples were collected from Lamb in Xinjiang area. A virus was isolated in MDBK cell line and primarily identified as the ORFV by electron microscope examination. Then the virus was inoculated to the sheep via skin scarification and the animal developed the typical signs of ORFV infection. Moreover, the B2L gene was amplified according to the ORFV sequence and clone into pET32a for expression in E.coli. SDS-PAGE analysis showed the recombinant B2L was about 60 ku and reacted positively with the ORFV positive serum identified by western blot. Furthermore, the phylogenetic analysis based on B2L gene indicated that the isolate was a high homology with the India 67/04 strain.

关 键 词：羊口疮病毒 B2L基因 原核表达 反应原性 

分 类 号：S852.65[农业科学—基础兽医学]