| 羊口疮病毒强弱毒株免疫相关基因的比较分析 |
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| 引用本文： | 张艳敏,丁学东,马园,张静,吉鹏华,王国华,潘相臣,刘冬冬,田普厚,石顺利,张七斤.羊口疮病毒强弱毒株免疫相关基因的比较分析[J].畜牧与饲料科学,2020,41(5):96-106. |
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| 作者姓名： | 张艳敏 丁学东 马园 张静 吉鹏华 王国华 潘相臣 刘冬冬 田普厚 石顺利 张七斤 |
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| 作者单位： | 内蒙古农业大学兽医学院,内蒙古呼和浩特010018;内蒙古自治区农牧业科学院,内蒙古呼和浩特010031;内蒙古必威安泰生物科技有限公司,内蒙古呼和浩特011500;内蒙古通辽市家畜繁育指导站,内蒙古通辽028000 |
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| 摘 要： | 通过对采集到的青岛市某发病羊场羊口疮病料（强毒）在山羊成纤维细胞上连续传90代（弱毒）后, 分别进行病毒基因组提取, 并对提取到的基因组分别命名为ORFV-QD和ORFV-RD。根据GenBank上发表的ORFV全基因组序列设计并合成3对特异性引物, 分别扩增ORFV-QD和ORFV-RD的B2L基因、F1L基因和VIR基因片段, 并将扩增得到的基因组片段分别克隆到pMD-19T载体上, 转化到DH5α感受态细胞中, 对重组质粒进行鉴定后送至测序公司进行测序, 用DNASTAR软件对测序结果进行拼接及3组基因之间核苷酸同源性比较分析, 将测序结果与NCBI上已公布的13组ORFV全基因组相应基因核苷酸序列进行同源性比对分析、构建系统进化树及氨基酸序列比对分析。结果表明, 3组基因与参考序列的B2L基因、F1L基因和VIR基因核苷酸同源性分别为92.1%~98.4%、96.1%~99.1%和94.6%~100%。将2株病毒基因组与参考序列进行氨基酸序列比较分析, 结果显示2株病毒基因组之间并没有较为明显的差异。
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| 关 键 词： | ORFV 免疫相关基因 系统进化树 同源性分析 |
| 收稿时间： | 2020-07-02 |
Comparative Analysis of Immune-related Genes Between Highly Virulent Strain and Attenuated Strain of Orf Virus (ORFV) |
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| ZHANG Yan-min,DING Xue-dong,MA Yuan,ZHANG Jing,JI Peng-hua,WANG Guo-hua,PAN Xiang-chen,LIU Dong-dong,TIAN Pu-hou,SHI Shun-li,ZHANG Qi-jin.Comparative Analysis of Immune-related Genes Between Highly Virulent Strain and Attenuated Strain of Orf Virus (ORFV)[J].Animal Husbandry and Feed Science,2020,41(5):96-106. |
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| Authors: | ZHANG Yan-min DING Xue-dong MA Yuan ZHANG Jing JI Peng-hua WANG Guo-hua PAN Xiang-chen LIU Dong-dong TIAN Pu-hou SHI Shun-li ZHANG Qi-jin |
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| Institution: | 1.College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China;2.Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences, Hohhot 010031, China;3.Inner Mongolia Bigvet Biotech Co., Ltd., Hohhot 011500, China;4.Livestock Breeding Guide Station of Tongliao City of Inner Mongolia, Tongliao 028000, China |
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| Abstract: | In this study, a highly virulent orf virus （ORFV） strain was isolated from the clinical samples of the diseased sheep. After 90 consecutive passages on goat fibroblasts, the obtained ORFV clinical strain was attenuated. The genomes of the highly virulent ORFV strain and the attenuated ORFV strain were extracted and designated as ORFV-QD and ORFV-RD, respectively. A total of 3 sets of specific primers were designed based on the whole genome sequence of ORFV published on GenBank and used to amplify the gene fragments of B2L, F1L and VIR in ORFV-QD and ORFV-RD. The obtained genomic fragments were cloned into pMD-19T vector and were subsequently transformed into DH5α competent cells. After identification, the positive recombinant plasmids were sequenced. The obtained target gene sequences were aligned, and the nucleotide sequence homology within the same gene between ORFV-QD and ORFV-RD was compared using DNASTAR software. Furthermore, the nucleotide sequence homology comparison, gene-based phylogenetic analysis and amino acid sequence homology comparison between the gene sequences obtained in this study and corresponding gene sequences from 13 ORFV whole genome sequences published on NCBI were carried out. The results showed that the nucleotide sequence homology of the B2L, F1L and VIR genes between the sequences obtained in this study and the reference sequences was 92.1%-98.4%, 96.1%-99.1%, and 94.6%-100%, respectively. The amino acid sequence homology analysis demonstrated that there were no obvious differences between ORFV-QD and ORFV-RD. |
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| Keywords: | ORFV immune-related genes phylogenetic tree homology analysis |
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