一株水貂博卡病毒全基因组扩增及序列分析  

Genome Amplification and Sequencing of the Mink Bocavirus

作　　者：于永乐 姚延珠 张传美 秦志华 张洪亮 杨瑞梅 段笑笑 单虎 YU Yongle;YAO Yanzhu;ZHANG Chuanmei;QIN Zhihua;ZHANG Hongliang;YANG Ruimei;DUAN Xiaoxiao;SHAN Hu(College of Veterinary Medicine,Qingdao Agricultural University,Qingdao 266109,China;College of Plant Health and Medicine,Qingdao Agricultural University,Qingdao 266109,China;Qingdao Animal Disease Prevention and Control Center,Qingdao 266000,China)

机构地区：[1]青岛农业大学动物医学院山东省预防兽医学重点实验室,青岛266109 [2]青岛农业大学植物医学学院,青岛266109 [3]青岛市动物疫病预防控制中心,青岛266000

出　　处：《病毒学报》2022年第2期439-447,共9页Chinese Journal of Virology

基　　金：在青高校服务青岛产业发展重点学科(兽医学)(项目号:青教办字2019-69);青岛农业大学高层次人才科研基金(项目号:663/1120015),题目:犬细小病毒VP2蛋白相关功能位点的研究。

摘　　要：本研究旨在研究水貂博卡病毒(Mink bocavirus,MBoV)在山东省的流行状况及基因特征。采用PCR对采自山东省境内水貂养殖场的85份水貂腹泻样品进行MBoV检测,挑选1份单阳性样品进行全基因扩增,使用MegAlign进行序列同源性比对分析,利用DNAMAN V6对基因组5’末端和3’末端回文结构进行预测,应用MEGA V6进行遗传进化分析。结果表明,所采集的腹泻样品中MBoV阳性率为3.5%(3/85),其中1份为MBoV单阳性,2份为MBoV与水貂肠炎病毒(Mink enteritis virus,MEV)双阳性。获得MBoV全基因序列1条(SDMBoV2020),全长5 252 bp;经分析,5’-和3’-UTR分别由98和145 bp短回文序列组成,具有典型的细小病毒末端的茎环样结构;基因组含有3个ORFs,分别编码非结构蛋白NS1、NP1以及结构蛋白VP1和VP2;SDMBoV2020与NCBI登录的MBoV参考毒株KU950356各编码基因的推导氨基酸序列同源性较高,为98.5%~99.0%;基于MBoV全基因序列构建的系统发育进化树也显示,SDMBoV2020和参考毒株KU950356株同处于MBoV 1型基因群分支上,亲缘关系较近。We investigated the prevalence and genetic characterization of the mink bocavirus(MBoV) in Shandong Province,China. Eighty-five mink samples were collected from mink farms. MBoV detection was done by polymerase chain reaction(PCR). The genomic DNA of the MBoV was amplified,cloned and sequenced from a single-positive sample. Multiple comparative analyses were conducted with the genomic sequences of the MBoV using MegAlign. The inverted terminal repeats of the 5’-untranslated region(UTR)and 3’-UTR were predicted by DNAMAN 6. Phylogenetic analyses were done by MEGA 6 based on the neighbor-joining method. The positivity rate of the MBoV was 3.5%(3/85),including one single-positive for the MBoV and two double-positives for MBoV and the mink enteritis virus(MEV). The whole genomic sequence was obtained by overlap PCR amplification and named as“SDMBoV2020”. The genomic sequence of SDMBoV2020 was submitted to GenBank under the accession number OK032607. The 5’-UTR and 3’-UTR consisted of 98-bp and 145-bp short palindromic sequences,respectively,with a typical stem loop-like structure at the end of the parvovirus;the genome contained three open reading frames encoding the non-structural proteins NS1 and NP1,as well as structural proteins VP1 and VP2,respectively. The deduced sequence of amino acids of SDMBoV2020 shared high homology with that of the MBoV reference strain KU950356,and showed 98.5% – 99.0% similarity. The phylogenetic tree constructed based on the full-length genomic sequences of the MBoV showed SDMBoV2020 to be located in the same branch as the reference strain KU950356,and to be closely related to it.

关 键 词：水貂博卡病毒 基因组扩增 进化分析 

分 类 号：S858.92[农业科学—临床兽医学]