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根据GenBank数据库公布的鱼类神经坏死病毒(nervousnecrosisvirus,NNV)同源序列设计了7对特异性引物,采用RT＿PCR方法扩增出目的片断,将PCR产物测序和分析。斜带石斑鱼Epinepheluscoioides神经坏死病毒(orange＿spottedNNV,OGNNV)基因组由两个片断(RNA1和RNA2)组成,RNA1由3103个核苷酸组成,含有一个开放阅读框,编码982个氨基酸;RNA2由1433个核苷酸组成,含有一个开放阅读框,编码338个氨基酸。OGNNV基因组与新加坡GGNNV(greasygrouperNNV)的基因组有高度的相似性。分析病毒的RNA2序列发现:OGNNV与DGNNV(dragongrouperNNV)、RGNNV(redspottedNNV)和GGNNV的亲缘关系很近,并且具有相同的中和位点;分析病毒的RNA1序列,发现在OGNNV的RNA1序列中同样可以找到依赖RNA的RNA聚合酶的6个模序(motif)。根据同源性比较和系统进化分析,OGNNV属于RGNNV血清型的成员。
Seven pairs of specific primers were designed according to the homologous sequences of nervousnecrosis virus (NNV) published in GenBank. The target fragment was amplified by RT-PCR and the PCR products were sequenced and analyzed. Epinepheluscoioides necrosis virus (orange_spottedNNV, OGNNV) genome consists of two fragments (RNA1 and RNA2), RNA1 consists of 3103 nucleotides, contains an open reading frame, encoding 982 amino acids; RNA2 consists of 1433 Nucleotide composition, containing an open reading frame, encodes 338 amino acids. The genome of OGNNV has a high degree of similarity with the genome of Singapore GGNNV (greasy grouperNNV). Analysis of the RNA2 sequence of the virus revealed that OGNNV has close genetic relationship with DGNNV (dragongrouperNNV), RGNNV (redspottedNNV) and GGNNV, and has the same neutralization site. Analyzing the RNA1 sequence of the virus, it was also found that in the RNA1 sequence of OGNNV, Find 6 motifs for RNA-dependent RNA polymerase. Based on homology comparison and phylogenetic analysis, OGNNV is a member of the RGNNV serotype.