蜜蜂残翼病毒中国DWV-JL1株衣壳蛋白基因序列分析  

Analysis on the Gene Sequence Encoded Capsid Protein of Chinese Deformed Wing Virus-JL1 Strain

作　　者：张健[1] 杨倩[1,2] 宋战昀[2] 郑言[2] 王向辉[2] 隋佳辰 王全凯[1,3] 王振国[2] 牟峻[2] 

机构地区：[1]吉林农业大学中药材学院,长春130118 [2]吉林出入境检验检疫局检验检疫技术中心,长春130062 [3]吉林省中韩动物科学研究院,长春130600

出　　处：《经济动物学报》2015年第2期109-113,124,共6页Journal of Economic Animal

基　　金：国家自然科学基金项目(31001065);国家质检总局科技计划项目(2013IK033);国际科技合作专项项目(2011DFA32900)

摘　　要：蜜蜂残翼病毒是引起蜜蜂残翼病的病原体。吉林出入境检验检疫局检验检疫技术中心将成功分离鉴定得到的中国首株蜜蜂残翼病毒毒株命名为“中国DWV—JL1株”，全长9826nt。本文主要研究中国DWV—JL1株核苷酸序列位置为2880～3792nt之间的片段，该段序列为编码衣壳蛋白的一部分，为相对保守序列。通过RT—PCR、克隆及质粒双酶切鉴定分析，得到一段大小约900bp的片段。通过BLAST、进化树及残基化分析，有96％-97％的同源性。氨基酸序列差异性仅为7％。该段序列与韩国株的亲缘关系最近，而与欧洲和美洲各国家获得的毒株序列亲缘关系较远，推测中国DWV—JL1株起源于亚洲。Wing deformities of honeybees have long been thought to associate with a virus, appropri- ately named deformed wing virus （DWV）. Our laboratory successfully isolated and identified a new DWV strain, named China DWV-JL1. It was consist of 9 826 nucleotide. Then the nucleotide se- quences encoded capsid protein was sequenced and was between 2 880 and 3 792 （913 nt）. Its gene sequence was relatively conservative. The sequence was cloned into plasmid and the recombinant plasmid was identified by double enzyme digestion analysis. And about 900 nt gene fragmen was obtained, which was equal to the theoretical size. By BLAST, the phylogenetic trees was constructed to determine the genetic relationship with the other strains according to the nucleotide sequence. The result showed that the sequence of the strain had 96%-97% homology with the other strains. The di- versity of amino acid sequence was only 7%. Overall, the Chinese DWV strain showed the closest genetic relationship with the Asian countries and a high divergence from the European and America countries, so it was suggested that the China DWV-JL1 strain came from Asia.

关 键 词：蜜蜂残翼病毒 RT-PCR 序列分析 进化树 衣壳蛋白 氨基酸序列 

分 类 号：S476.13[农业科学—农业昆虫与害虫防治]