福建农业科技2025，Vol.56Issue(12)：10-15,6.DOI:10.13651/j.cnki.fjnykj.2025.12.002
石斑鱼神经坏死病毒衣壳蛋白多克隆抗体的制备
Preparation of Polyclonal Antibody Against the Capsid Protein of Grouper Nervous Necrosis Virus
摘要
Abstract
Grouper nervous necrosis virus(GNNV)infects the nervous system of juvenile fish,with the mortality rates as high as 90%-100%,which was a bottleneck restricting the development of grouper aquaculture.The preparation of polyclonal antibodies against GNNV is essential for the in-depth study of viral protein functions and the development of related detection methods.In this study,an expression vector containing the protruding domain(P-domain)of the GNNV capsid protein,designated GNNV-P-pET-15b,was constructed and transformed into Escherichia coli BL21(DE3)for the prokaryotic expression.The purified GNNV-P protein was used as an antigen to immunize New Zealand white rabbits,generating the rabbit-derived polyclonal antiserum.The antiserum titer was determined by indirect ELISA,and its specificity was analyzed by Western blot.The results showed that:the 15 kDa soluble recombinant GNNV-P protein was successfully obtained by prokaryotic expression of GNNV-P-pET-15 b vector,and then further used as an antigen to immunize New Zealand white rabbits to prepare the rabbit-derived polyclonal antiserum against GNNV-P,with the titer of antiserum exceeding 1∶150 000 by using the method of indirect ELISA.The Westernblot showed that GNNV-P polyclonal antibody had high specificity.The results provided key biological materials for the subsequent study of GNNV infection mechanism and the development of related immunological detection methods.关键词
石斑鱼神经坏死病毒/衣壳蛋白/突出结构域/多克隆抗体Key words
Grouper nervous necrosis virus/Capsid protein/Protruding domain/Polyclonal antibody分类
农业科技引用本文复制引用
王焌翔,江金连,曾思瑶,陈美莲,吴雅玲,林琳..石斑鱼神经坏死病毒衣壳蛋白多克隆抗体的制备[J].福建农业科技,2025,56(12):10-15,6.基金项目
福建省科技计划重大专项专题(2022NZ033021) （2022NZ033021）
福建省自然科学基金项目(2023J011576) （2023J011576）
福建省促进海洋与渔业产业高质量发展专项资金项目(FJHYF-L-2025-16) （FJHYF-L-2025-16）
福州海洋研究院"揭榜挂帅"科技项目(2022F15、2024F12、2025F15) （2022F15、2024F12、2025F15）
闽江学院校长基金(103952024046). （103952024046）
