西方蜜蜂囊状幼虫病病毒云南毒株的多聚蛋白基因扩增及序列分析  

Amplification and sequence analysis of polyprotein gene of sacbrood virus Yunnan strain from Apis mellifera

作　　者：曹兰[1] 沈克飞[1] 王瑞生[1] 张邑帆[1] 高丽娇[1] 张祖芸[2] 曹联飞[3] CAO Lan;SHEN Ke-fei;WANG Rui-sheng;ZHANG Yi-fan;GAO Li-jiao;ZHANG Zu-yun;CAO Lian-fei(Chongqing Academy of Animal Sciences,Chongqing 402460,China)

机构地区：[1]重庆市畜牧科学院,重庆402460 [2]云南省农业科学院,云南红河661101 [3]浙江省农业科学院,浙江杭州310021

出　　处：《中国生物制品学杂志》2021年第6期688-693,共6页Chinese Journal of Biologicals

基　　金：国家自然科学基金青年科学基金(31602014);国家现代农业(蜜蜂)产业技术体系建设专项(CARS-44-SYZ14);重庆市自然科学基金(cstc2020jcyj-msxmX1047);浙江省农业(畜禽)新品种选育重大科技专项(2016C02054-11)。

摘　　要：目的扩增云南地区西方蜜蜂(Apis mellifera)自然感染囊状幼虫病病毒(sacbrood virus,SBV)的多聚蛋白(polyprotein)基因,并进行序列分析。方法采用RT-PCR方法分别从云南3份感染SBV的西方蜜蜂幼虫样本中扩增SBV的多聚蛋白基因,进行序列测定、拼接、序列比对、基因重组及分子进化分析。结果通过序列拼接得到3条云南西方蜜蜂的SBV(AmSBV-YN)序列,占SBV基因组长度的97%,包含完整的多聚蛋白基因。3条AmSBV-YN序列间的同源性大于99%,与GenBank上已知SBV序列的同源性为91%~93%。AmSBV-YN与韩国西方蜜蜂的SBV(AmSBV-Kor)_KP296800可能在5′端发生重组。多聚蛋白基因构建的进化树显示,AmSBV-YN单独构成云南基因型,在RNA依赖的RNA聚合酶(RNA-dependent RNA polymerase,RdRp)基因进化树中显示,AmSBV-YN与其他感染中国西方蜜蜂的SBV(AmSBV-Chn)聚集成云南基因型。结论成功扩增出SBV云南毒株的多聚蛋白基AmSBV-YN,SBV具有丰富的遗传多样性,AmSBV-YN可能是感染我国西方蜜蜂SBV的代表株。Objective To amplify and analyze the sequence of polyprotein gene of sacbrood virus(SBV)Yunnan strain(AmSBV-YN)from naturally infected Apis mellifera.Methods Polyprotein gene was amplified by RT-PCR from three larva samples of A.mellifera,then sequenced,spliced and subjected to sequence alignment,gene recombination and molecular evolution analysis.Results Three gene sequences of SBV(AmSBV-YN)were obtained by sequence splicing,which covered 97%of full-length of the SBV genome,and contained complete polyprotein gene.The homology of three AmSBV-YN gene sequences was more than 99%,while those to the known SBV gene sequences in GenBank were 91%~93%.AmSBV-YN might be recombined with AmSBVKor_KP296800 at the 5′-terminus.The phylogenetic tree based on polyprotein gene showed that AmSBV-YN consisted of Yunnan genotype alone.However,the phylogenetic tree based on RNA dependent RNA polymerase(RdRp)gene showed that AmSBV-YN and other SBV infecting A.mellifera in China(AmSBV-Chn)were clustered into Yunnan genotype.Conclusion The polyprotein gene of SBV Yunnan strain(AmSBVYN)was successfully amplified.Sequence analysis suggested abundant genetic diversity of SBV,and AmSBV-YN might be the representative strain of AmSBV-Chn.

关 键 词：蜜蜂 囊状幼虫病病毒 多聚蛋白基因 同源性 进化树 

分 类 号：Q65.8[生物学—生物物理学]