中国明对虾肝胰腺细小病毒全基因组的克隆及序列分析  

Genome cloning and sequence analysis of hepatopancreatic parvovirus(HPV) isolated from Fenneropenaeus chinensis in China

作　　者：张峥[1,2] 黄倢[1,2] 高强[1,3] 成君军[1] 王明森[1] 

机构地区：[1]农业部海洋渔业资源可持续利用重点开放实验室,中国水产科学研究院黄海水产研究所,山东青岛266071 [2]上海海洋大学,上海201306 [3]中国科学院海洋研究所,山东青岛266071

出　　处：《中国水产科学》2011年第1期59-65,共7页Journal of Fishery Sciences of China

基　　金：国家863计划项目(2006AA100306);国家重点基础研究发展计划项目(2006CB101801);农业部公益性行业(农业)科研专项(200803012);国家虾现代产业技术体系(nycytx-46)岗位专家经费项目

摘　　要：中国明对虾肝胰腺细小病毒是一种单链、线性DNA病毒。本研究利用DNA末端加尾和嵌套PCR首次完成了对虾肝胰腺细小病毒(hepatopancreatic parvovirus,HPV)中国分离株(HPV-Pc)全基因组序列的测定,研究结果表明,HPV-Pc株(GenBank登录号GU371276)的全基因组由6 085个核苷酸组成,包含3个开放阅读框(ORF)(分别代表ns1、ns2和cp基因)和2个非编码区。ORF1、ORF2和ORF3分别编码由427、578和821个氨基酸组成的蛋白,分子量分别为50.4 kD、68.2 kD和92 kD。分析HPV-Pc基因组结构,发现该基因组没有末端反向重复序列(ITR),cp基因编码的CP区有HPV特异的"GAA"正向重复序列,及富含甘氨酸的上游和下游不同位置存在精氯酸残基,可造成蛋白裂解。不同地区、不同对虾HPV基因组有较大差异,HPV-Pc株基因组与其他已知HPV序列同源性在78%～91%之间,差异遍布整个序列中。将HPV-Pc各基因片段与已报道的其他地区HPV相应序列进行比较,结果表明,HPV-Pc各基因片段与AY008257(韩国株)同源性最高。与已知HPV代表株的NS2、NS1和CP蛋白氨基酸序列同源性比较也显示,HPV-Pc株与韩国株同源性最高。这些特征表明HPV-Pc属于细小病毒科的一个新分离株。Hepatopancreatic parvovirus（HPV） is a single-stranded DNA virus.A HPV strain named as HPV-Pc was isolated from infected Fenneropenaeus chinensisin from Tangshan of Hebei Province,China.In this study,full-length genomic sequence of HPV-Pc（GenBank Accession no.GU371276） was determined.It consists of 6 085 nucleotides and contains three open reading frames which encode for non-structural protein（NS1 and NS 2） and nucleocapsid protein（CP）.The ORF1,ORF2 and ORF3 encode for proteins of 427,578 and 821 amino acids,whose molecular weight are 50.4 kD,68.2 kD and 92 kD,respectively.No inverted terminal repeat（ITR） was de-tected in 5′ terminal end of the genome.The ＂R＂ motif rich in ＂G＂ in the region of CP was detected,which may result in cleaving of CP protein.The ＂GAA＂ direct repeats were found in the region of CP,which was specific to HPV.Genomes of HPV in different shrimp and from different region were different.All of the acquired fragments were spliced by biological software.Full-length HPV genome shows similarity of 78%-91% with other known HPV genome,and the differences existed throughout its sequence.Comparison analysis between HPV-Pc and other HPV strains submitted to GenBank showed that the HPV-Pc had a larger cp gene and longer genome size.The nucleotide homology of HPV-Pc with AY008257（Korea strain） was the highest.Comparison analysis in NS2,NS1 and CP protein sequence also showed the highest homology existing between HPV-Pc and AY008257.These results suggest that HPV-Pc is a new isolate in Parvoviridae.This is the first nucleotide sequence of F.chinensis.

关 键 词：肝胰腺细小病毒(HPV) 中国明对虾 基因组序列 

分 类 号：S94[农业科学—水产养殖]