Hyink O, Graves S, Fairbairn F M, Ward V K
Department of Microbiology, University of Otago, Dunedin, New Zealand.
J Gen Virol. 1998 Nov;79 ( Pt 11):2853-62. doi: 10.1099/0022-1317-79-11-2853.
The light brown apple moth, Epiphyas postvittana, is a major insect pest of a variety of fruit crops grown in New Zealand and we are studying a nucleopolyhedrovirus, EppoNPV, isolated from this insect. Restriction endonuclease analysis of EppoNPV DNA shows that this is a single strain of virus with a genome size of approximately 119 kbp and a complete library of the EppoNPV genome has been cloned. A strategy of single-stranded sequencing of the termini of REN fragment clones was employed to map the virus genome. Sequence homologies to NPV gene sequences present in the GenBank database allowed a nearly complete restriction map of the EppoNPV genome to be constructed. The mapping was completed with Southern blotting and restriction analysis. Fifty-five open reading frames (ORFs) with similarity to genes from other NPVs have been identified and placement of these on the restriction map shows that EppoNPV has a nearly identical genome organization to Orgyia pseudotsugata (Op)MNPV. The polyhedrin gene of EppoNPV has been fully sequenced and an ORF of 738 bp encodes a predicted protein of 28.8 kDa. The conserved 12 bp consensus sequence typical of very late baculovirus gene promoters, AATAAGTAATTT, has been located upstream of the ATG initiation codon. An ORF located downstream of the polyhedrin gene shows homology to the 1629-capsid protein from OpMNPV. Phylogenetic comparison to polyhedrin gene sequences from 23 other NPVs shows EppoNPV to be a group I NPV closely related to OpMNPV.
浅褐苹果蛾（Epiphyas postvittana）是新西兰种植的多种水果作物的主要害虫，我们正在研究从这种昆虫中分离出的一种核型多角体病毒（EppoNPV）。对EppoNPV DNA进行的限制性内切酶分析表明，这是一种单一病毒株，基因组大小约为119 kbp，并且已经克隆了EppoNPV基因组的完整文库。采用对REN片段克隆末端进行单链测序的策略来绘制病毒基因组图谱。与GenBank数据库中存在的NPV基因序列的序列同源性使得能够构建EppoNPV基因组的近乎完整的限制性图谱。通过Southern印迹和限制性分析完成了图谱绘制。已鉴定出55个与其他NPV基因相似的开放阅读框（ORF），将这些ORF定位在限制性图谱上表明EppoNPV与云杉毒蛾（Op）MNPV具有几乎相同的基因组结构。EppoNPV的多角体蛋白基因已被完全测序，一个738 bp的ORF编码一个预测的28.8 kDa蛋白质。杆状病毒极晚期基因启动子典型的保守12 bp共有序列AATAAGTAATTT位于ATG起始密码子上游。位于多角体蛋白基因下游的一个ORF与OpMNPV的1629衣壳蛋白具有同源性。与其他23种NPV的多角体蛋白基因序列进行系统发育比较表明，EppoNPV是与OpMNPV密切相关的I组NPV。
