赤点石斑鱼神经坏死病毒主衣壳重组蛋白多克隆抗体的制备

把赤点石斑鱼（Epinephelus akaara）神经坏死病毒（RGNNV）主衣壳蛋白（MCP）基因的重组表达质粒载体pRSETA-MCP转化至大肠杆菌（Estherichia coli）BL21（DE3）,经IPTG诱导表达,SDS-PAGE显示表达的重组蛋白主要以不可溶的包涵体形式存在,分子量约44.5kD。通过Ni-NTA-Agarose亲和层析柱纯化,经分析纯度达90%,之后免疫新西兰兔制备抗血清,ELISA效价达1：12800以上。Western-blot分析结果显示,该血清与表达的重组蛋白有较强反应,说明通过原核表达的重组蛋白具有良好的免疫原性。

Preparation of polyclonal antibody for major capsid protein (MCP) of RGNNV

We transformed the recombinant expression plasmid vector pRSET A-MCP of major capsid protein gene（MCP） of red-spotted grouper（Epinephelus akaara）nervous necrosis virus（RGNNV） into Estherichia coli BL21（DE3） and induced the bacterial cultures with IPTG.SDS-PAGE analysis shows that the recombinant protein with the molecular weight of 44.5 kD is presented in the form of inclusion body.The recombinant protein is purified by affinity chromatography on Ni-NTA-Agarose column with 90% purity and then is used to immunize the New Zealand rabbit.ELISA result demonstrates that the titer of antiserum is over 1：12 800.Moreover,the Western-blot result reveals that the antiserum has a strong reaction with the recombinant protein expressed from E.coli BL21,and the prokaryotic expression of recombinant protein is proved to have good immunogenicity.