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Mmapping the neutralizing epitope on the coat protein of fish nodavirus.

定位鱼类诺达病毒外壳蛋白上的中和表位。

基本信息

批准号：
09660201
负责人：
NISHIZAWA Toyohiko
金额：
$ 1.98万
依托单位：
HIROSHIMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

The striped jack nervous necrosis virus (SJNNV), the causative agent of viral nervous necrosis in marine fish, is a members of fish nodaviruses. The fish nodavirus is an isometric virus consisting of a single type of coat protein and two positive-sense RNA molecules, and is divided into four different genotypes (SJ, TP, BF and RG types) based on the nucleotide sequence of the coat protein gene (RNA2). In this study, two different regions of the coat protein gene of SJNNV and other genotypes of fish nodaviruses were cloned into expression vector plasmids for analysis of the serological relationships and mapping the neutralizing epitope on SJNNV.The four different genotypes of nodaviruses were confirmed to be distinguishable from each other easily by the hybridization with DIG labeled DNA probes. Furthermore, by western blot analysis with A/S SJNNV and MAbs having neutralizing activities against SJNNV, it was revealed that SJNNV and other fish nodaviruses shared a significant number of antigenic determinants with other genotype viruses, although SJNNV was serologically distinguishable from them. And it was suggested that the antigenic determinant of SJNNV for the neutralizing MAbs was a liner epitope, which consisted of a repeated amino acid sequence at the coat protein. It was considered that one of the neutralizing epitopes of SJNNV was PAN (aa 254-256). The other genotypes of fishnodaviruses, TP, BF and RG types, have a PPG, PEG and/or PDG at the same amino acid position as SJNNV coat protein, thus it was considered that the four different genotypes of fish nodaviruses were also serologically different from each other.

条纹杰克神经坏死病毒（striped jack nervous necrosis virus，SJNNV）是鱼类诺达病毒属的成员，是引起海水鱼类病毒性神经坏死的病原。鱼类诺达病毒是由单一类型的外壳蛋白和两个正义RNA分子组成的等轴病毒，根据外壳蛋白基因（RNA 2）的核苷酸序列可分为四种不同的基因型（SJ、TP、BF和RG型）。本研究将SJNNV外壳蛋白基因的两个不同区域与其他基因型的鱼诺达病毒克隆到表达载体质粒中，分析了SJNNV的血清学关系，并定位了SJNNV的中和表位。此外，通过用A/S SJNNV和具有抗SJNNV中和活性的单克隆抗体进行蛋白质印迹分析，发现SJNNV和其他鱼诺达病毒与其他基因型病毒共享大量抗原决定簇，尽管SJNNV与它们在血清学上是可区分的。提示SJNNV的中和性单克隆抗体的抗原决定簇为线性表位，由外壳蛋白的重复氨基酸序列组成。认为SJNNV的中和表位之一是PAN（aa 254-256）。鱼诺达病毒的其他基因型TP、BF和RG型在与SJNNV外壳蛋白相同的氨基酸位置具有PPG、PEG和/或PDG，因此认为鱼诺达病毒的四种不同基因型在血清学上也彼此不同。