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根据鲤疱疹病毒II型(Cyprinid herpesvirus II,CyHV-2)ORF 4基因序列(GenBank：JQ815364．1)设计特异性引物,PCR扩增得到ORF 4基因编码框全长序列1041 bp,将其克隆至原核表达载体pET-32a (+)中,构建了重组原核表达载体pET-32a-ORF 4。将pET-32a-ORF 4重组载体转化大肠杆菌BL21(DE3),经IPTG诱导得到融合表达的重组蛋白,融合表达的重组蛋白主要以包涵体的形式存在,其分子质量约为57 ku,与预期大小一致。将纯化的重组蛋白免疫日本大耳兔,制备了多克隆抗体, ELISA检测抗体效价大于1∶50000, Western blot检测显示该抗体可以特异性识别重组蛋白。间接免疫荧光检测结果表明：该多克隆抗体可与由CyHV-2感染引起细胞病变的异育银鲫脑组织细胞( GICB)发生特异性的结合。“,”The specific primer pairs were designed according to the ORF 4 gene sequence of Cyprinid herpesvirus II ( Cy-HV-2) and the 1041bp full-length sequence of ORF4 gene coding frame was amplified by PCR, and then the full-length CyHV-2 ORF 4 gene was cloned into prokaryotic expression vector pET-32a( +) to construct the recombinant ex-pression vector pET-32a-ORF 4. The recombinant expression vector pET-32a-ORF 4 was successfully expressed in E. coli BL21 (DE3) as a fusion protein with a size of 57 ku after induction by isopropyl-β-D-thiogalactoside (IPTG). The recombinant protein was presented mainly in inclusion body. The purified recombinant proteins were used to immune Japanese white rabbit to produce polyclonal antibody. Titer of polyclonal antibodies was above 1∶50 000. Western blot test demonstrated that the recombinant protein was recognized by the polyclonal antibody.