作　　者： ; ; ; ; ;

机构地区： 中山大学生命科学学院水生经济动物繁殖营养和病害控制国家重点学科专业实验室

出　　处： 《水产学报》 2004年第2期183-188,共6页

摘　　要： 从患病毒性神经坏死病的斜带石斑鱼(Epinepheluscoioids)的头部提取总RNA,根据已发表的神经坏死病毒外壳蛋白基因设计引物进行RT PCR扩增,得到预期大小的基因片段。将此基因片段转入pET载体进行序列测定和分析,结果表明:编码斜带石斑神经坏死病毒(Orange spottedgroupernervousnecrosisvirus,OGNNV)外壳蛋白基因的阅读框核苷酸数为1017bp,编码338个氨基酸;基因的核苷酸序列与野田村病毒科(Nodaviridae)的几种病毒的外壳蛋白基因序列比较结果显示,该病毒与β野田村病毒属(Betanodavirus)中的赤点石斑神经坏死病毒(red spottedgroupernervousnecrosisvirus,RGNNV)的同源性最高(99%),说明该病毒株是RGNNV血清型的成员。 The nodaviridae, which include two genera: alphanodavirus and betanodavirus, are a family of small, isometric and non-enveloped RNA viruses.Alphanodaviruses primarily infect insects and betanodaviruses infect fish. Their genomes, which are among the smallest known for animal viruses, consist of two co-encapsidated positive-sense RNA segments: RNA1 encodes the RNA-dependent RNA polymerase (RdRp) which replicates the viral genomes, while RNA2 encodes a precursor to the coat protein. Based on the coat protein sequence, betanodaviruses are categorized into four different genotypes: TPNNV(tiger puffer nervous necrosis virus), SJNNV(striped jack nervous necrosis virus), BFNNV(barfin flounder nervous necrosis virus),RGNNV(red-spotted grouper nervous necrosis virus).Orange-spotted grouper (Epinephelus coioides) nervous necrosis virus (OGNNV) is the causative agent of viral nervous necrosis (VNN) in larval and juvenal orange-spotted grouper fish. In this study, total RNA was extracted from the head of orange-spotted grouper infected with VNN. A pair of primers were designed according to the homogenus gene sequences, which have been submitted to GeneBank, from other fish nervous necrosis virus (NNV). The forward (sense) primer contained SacⅠsite and the reverse(antisense) primer contained HindⅢ sites. One target fragment was amplified by means of reverse-transcriptase polymerase chain reaction (RT-PCR). After digestion with SacⅠ and Hand Ⅲ, the RT-PCR products were ligated into the pET32 plasmid vector and transformed into E.coli BL21. The RT-PCR product and recombinant plasmid were sequenced , respectively. The coat protein gene of OGNNV was 1017 bases, encoded a protein of 338 amino acid with molecular mass of 37059.88 da, and with isoelectric point (IP) of 8.87. The sequence of the coat protein gene from OGNNV was submitted to GeneBank and the accession number was AF534998.The nucleotide sequence of the coat protein gene from OGNNV was aligned with four betanodaviruses: SJNNV,TPNNV,RGNNV,BFNNV and one alphanodav

关 键 词： 斜带石斑鱼 神经坏死病毒 外壳蛋白 基因克隆 序列分析 野田村病毒科 野田村病毒属

领　　域： [农业科学] [农业科学]