羊口疮病毒上海流行株的分离鉴定及其保护性抗原基因遗传进化分析  

Isolation,identification and protective antigen genes phylogenetic analysis of Orf virus epidemic strains in Shanghai

作　　者：陈晓 李丽 葛雷 李垚[4] 戴建军[2,3] 王书全 张树山[2,3] 吴彩凤[2,3] 张德福[2,3] CHEN Xiao;LI Li;GE Le;LI Yao;DAI Jian-jun;WANG Shu-quan;ZHANG Shu-shan;WU Cai-feng;ZHANG De-fu(College of Animal Medical University, Jinzhou,Liaoning 121001, China;In nary Medicine, Shanghai Academy of Agricultural Science Husbandry and Veterinary, J inzhou stitute of Animal Science and Veteri- s, Shanghai 201106, China;Division of Animal Genetic Engineering, Shanghai Municipal Key Laboratory of Agri-genetics and Breeding, Shanghai 201106, China;College of Medicine, Shanghai J iaotong University, Shang- hai 200025,China)

机构地区：[1]锦州医科大学畜牧兽医学院,辽宁锦州121001 [2]上海市农业科学院畜牧兽医研究所,上海201106 [3]上海市农业遗传育种重点实验室动物遗传工程研究室,上海201106 [4]上海交通大学医学院,上海200025

出　　处：《中国兽医学报》2018年第4期655-661,共7页Chinese Journal of Veterinary Science

基　　金：上海市科委实验动物专项基金资助项目(15140900300,17140900100);上海市农委青年人才计划资助项目[沪农青字(2015)第1-36号];上海市农业科学院攀高计划资助项目(PG171)

摘　　要：为了解上海地区羊口疮病毒（ORFV）流行毒株的分子生物学特性与遗传变异情况，采集疑似0RFV感染的羔羊唇部结痂组织，应用PCR、MDBK细胞分离培养、电子显微镜观察、间接免疫荧光技术等方法，进行分离鉴定，并对这2个分离毒株保护性抗原基因F1L、B2L进行克隆与序列分析。结果表明，成功分离到2株ORFV，将其分别命名为ORFuF416、ORFV-F429。其中，2株分离株F1L基因与参考毒株核苷酸序列同源性分别为95．2％～99．4％，推导氨基酸序列与参考毒株氨基酸序列同源性分别为94．4％～99．4％，遗传进化树分析均显示2株均属于同一分支，且与福建株FJ—YX（KC568410）亲缘关系最近。但与相关参考毒株相比，分离株FIL基因编码的氨基酸在44～66位不存在核苷酸的缺失，表明上海流行株与FJ—YX株存在一定的变异。2株分离株B2L基因与参考毒株核苷酸序列同源性为96．7％～99．6％，氨基酸序列同源性为94．7％～98．9％，且在遗传进化树上均与广西株GX—YB（JQ904793）有较近的亲缘关系，但与中国疫苗株（JQ904789）亲缘关系较远。表明从上海地区分离得到的2个毒株与流行于中国南方地区的0RFV毒株有较近的亲缘关系，但分离株的FIL基因和B2L基因存在一定变异。In order to understand the molecular biological characteristics and genetic evolution of the epidemic strains of the Orf virus（ORFV） in Shanghai,the scabs of lambs infected with suspec- ted ORFV were collected and analyzed by PCR, MDBK cells, electron microscopy,indirect immu- nofluorescence technology and other experiments for isolation and identification of virus, and the protective antigen gene F1L,B2L of two isolates were cloned and sequenced. The results showed that two strains of ORFV were successfully isolated and named ORFV-F416 and ORFV-F429 re- spectively. Their nucleotide sequence homology of the F1L gene were 95.2%-99.4% with the ref- erence strains,and the amino acid sequence of the deduced was 94.4%-99. 4% ,and the genetic phylogenetic tree analysis the two strains belonged to the same branch,and was closely related to Fujian FJ-YX strain（KC568410）. However, there was no nucleotide deletion in the 44-66 amino acid of the FIL gene compared with the related reference strain,which indicated that there was a certain variation between the Shanghai epidemic strain and the FJ-YX strain. The homology of the nucleotide sequence of two isolates of the B2L gene was 96.7%-99.6% with the reference strain, the amino acid sequence homology was 94. 7%-98. 9%, and it was close to the Guangxi GX-YB （JQ904793）,but was far from the China vaccine strain （JQ904789 ）. The two isolates from the Shanghai area are closely related to the ORFV strains prevalent in the southern China, but the FIL and B2L genes of the isolates are somewhat different.

关 键 词：ORFV 分离鉴定 F1L基因 B2L基因 进化分析 

分 类 号：S852.65[农业科学—基础兽医学] R535[农业科学—兽医学]