黑蜂王台病毒中国BQCV-JL1株的分离鉴定及全基因组序列分析  

Isolation,Identification and Analysis of the Complete Genome Sequence of Black Queen Cell Virus Strain China-JL1

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

机构地区：[1]吉林出入境检验检疫局检验检疫技术中心,长春130062 [2]吉林农业大学,长春130118 [3]长春生物制品研究所有限责任公司,长春130062

出　　处：《病毒学报》2015年第2期114-123,共10页Chinese Journal of Virology

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)

摘　　要：收集吉林蜂场的蜜蜂蜂蛹病料的RNA作为扩增模板,依据GenBank公布的黑蜂王台病毒(Black queen cell virus,BQCV)的全基因组序列,自行设计了10对引物,运用RT-PCR首次获得中国BQCV毒株的全基因组序列,命名为中国BQCV-JL1株。其全基因组序列由8 358个碱基组成,与GenBank公布的其他6株BQCV毒株的全基因组序列相比,同源性为86%~93%。中国BQCV-JL1株ORF 1的位置为546nt^4 676nt(4 131nt),ORF 2位于5 750nt^8 203nt,两个ORFs之间存在一段长为543nt(4 891nt^5 433nt)的ORF3。中国BQCV-JL1株第一个基因功能区ORF1′的位置为546nt^5 429nt,包含ORF 1和ORF 3。在ORF2之前存在内部核糖体进入位点(IRES),其末尾3碱基为CCU(5 642nt^5 644nt),为促进ORF 2翻译的起始密码子,中国BQCV-JL1株第二个基因功能区ORF2′的位置为5 642nt^8 203nt。中国BQCV-JL1株与Hungary 10(EF517515)同源性最高(93%),且分析核苷酸序列和氨基酸序列,与South Korea(JX149531)株最接近,表明中国BQCV-JL1株与South Korea(JX149531)株均有可能来自欧洲。中国BQCV-JL1株与其他6株全基因组序列在ORF位置划分上存在差异,其原因是部分位置发生基因突变。Honeybee pupae were collected from Jilin apiaries and RNA was extracted for use as a template for amplification.Based on the complete genome sequences of black queen cell virus（BQCV）published on GenBank,we designed 10 pairs of primers to amplify genes by reverse transcription-polymerase chain reaction（RT-PCR）.Using this approach,we have obtained the first complete genome sequence of a BQCV isolate in China.The genome of the isolated strain,named BQCV-JL1,is composed of 8358 nucleotides and shares between 86% and 93% homology with the complete genome sequences of the other six BQCV strains published on GenBank.ORF 1of BQCV-JL1 is positioned between nucleotides（nt）546and 4 676（4 131nt）,while ORF 2is located between nt 5 750 and 8 203.Between the two ORFs of BQCV-JL1 there is a short ORF,called ORF 3,between nt 4 891 and 5 433（543nt）.The first functional gene expression domain of the BQCV-JL1 strain is positioned between nt 546 and 5 429,encompassing both ORF1 and ORF 3.There is an internal ribosome entry site（IRES）located before ORF 2,the last three bases of which are CCU（nt 5 642-5 644）.These bases act as an initiation codon facilitating the translation of ORF 2.The second functional gene expression domain of the BQCV-JL1 strain is located between nt positions 5 642 and 8 203.The BQCV-JL1 strain was found to share high sequence identity（93%）with the Hungary 10 genotype at the whole-genome level and analysis of the nucleotide and amino acid sequences revealed that the BQCV-JL1 strain also shows close genetic relationships with the South Korea strain,suggesting that both the BQCV-JL1 and South Korea strains may have migrated from European countries.BQCV-JL1 strain was different from the other 6strains in dividing the nucleotides positions of ORF,which was because of the gene mutation.

关 键 词：BQCV 全基因组序列 RT-PCR 进化树分析 RDRP 

分 类 号：S476.13[农业科学—农业昆虫与害虫防治]