羊源与骆驼源羊口疮病毒免疫相关基因的比较分析

试验旨在探究羊口疮病毒（Orf virus，ORFV）在感染不同物种后其免疫相关基因所表现出的差异变化。对采集到的羊和骆驼ORFV病料进行病毒基因组提取，分别命名为ORFV-Y和ORFV-LT。根据GenBank中ORFV全基因组序列（登录号：KF234407.1）设计并合成3对特异性引物，分别扩增ORFV-Y和ORFV-LT的B2L、F1L和VIR基因片段，并将扩增得到的基因片段分别克隆到pMD19-T载体上，转化大肠杆菌DH5α感受态细胞，对重组质粒进行鉴定，选取阳性克隆质粒进行测序，用DNAStar软件对所获序列进行拼接后，与NCBI上已公布的13组ORFV全基因组相应基因序列进行同源性比对、氨基酸序列分析和系统进化树构建。结果显示，两组基因组与参考序列的B2L、F1L和VIR基因核苷酸同源性分别为92.8%~99.2%、95.7%~99.5%和77.6%~100%。将两组基因组与参考序列进行氨基酸序列比对分析后发现，两组基因组之间的免疫相关基因均表现出较为明显的差异，且F1L基因有一定的规律可循。对B2L、F1L和VIR基因进行系统进化树构建分析后发现，ORFV-Y与中国福建山羊株亲缘关系较近，而ORFV-LT与参考毒株进化关系均较远，且单独成为一个分支。综上，ORFV在感染羊和骆驼时其免疫相关基因出现了较为明显的差异，为深入探究ORFV感染不同物种其基因序列发生的变化及未来针对不同物种的疫苗研制提供参考。

Comparison Analysis of Immune Related Gene of Orf Virus in Sheep and Camels

The purpose of this study was to investigate the variation of Orf virus (ORFV) immune related genes after infection with different species.The ORFV genomes of sheep and camel were extracted and named ORFV-Y and ORFV-LT,respectively.Based on ORFV genome sequence published in GenBank (accession No.:KF234407.1),three pairs of specific primers were designed and synthesized to amplify the B2L,F1L and VIR gene fragments of ORFV-Y and ORFV-LT,respectively,and the amplified fragments were cloned into pMD19-T vector,transformed into E.coli DH5α competent cells.The recombinant plasmid was identified,and positive clones were selected for sequencing,DNAStar software was used to analyze the homology,amino acid sequence and phylogenetic tree of 13 ORFV genome sequences published on NCBI.The results showed that the nucleotide homology of B2L,F1L and VIR genes were 92.8% to 99.2%,95.7% to 99.5% and 77.6% to 100%,respectively.After comparing the amino acid sequence between the two genomes and the reference sequence,it was found that there were obvious differences in the immune related genes between the two genomes,and F1L gene had some rules to follow.The phylogenetic analysis of B2L,F1L and VIR genes showed that ORFV-Y was closely related to the Chinese Fujian goat strain,while ORFV-LT was far from the reference strains,and was a separate branch.The results showed that ORFV had obvious difference in immune related genes between sheep and camels,it provided a reference basis for further research on the changes of ORFV gene sequences in different species and the development of vaccines for different species in the future.