鲤疱疹病毒3型ORF57基因工程亚单位疫苗制备及免疫保护研究

旨在研究鲤疱疹病毒3型(Cyprinid herpesvirus 3,CyHV-3)囊膜蛋白ORF57作为疫苗候选抗原的可能性。根据已经发表的鲤疱疹病毒3型全基因组(GenBank登录号为 DQ657948.1),设计特异性引物,扩增、克隆ORF57序列并进行生物信息学分析。构建重组原核表达载体pET-32a(+)-CyHV-3-ORF57,大肠埃希菌体外表达获得重组蛋白ORF57。此外,对ORF57亚单位疫苗的免疫保护性进行研究。鲤鱼免疫4周后CyHV-3攻毒具有75%的存活率,ORF57亚单位疫苗组鲤鱼的存活率显著高于PBS和pET-32a(+)组。实验结果证明,重组ORF57亚单位疫苗具有较强的免疫保护性,为免疫学临床的应用和CyHV-3疫苗的开发提供了实验基础。

Preparation and immunoprotection of ORF57 subunit vaccine of Cyprinid herpesvirus 3

The aim of this study was to study the possibility of ORF57, an important capsule protein of Cyprinid herpesvirus 3, as a vaccine candidate antigen. According to the published genome sequence of Cyprinid herpesvirus 3 (GenBank number: DQ657948.1), specific primers were designed to amplify and clone the ORF57 sequence, and bioinformatics analysis was performed. The recombinant prokaryotic expression vector pET-32a(+) -CyHV-3-ORF57 was constructed, and the recombinant protein ORF57 was expressed by E. coli in vitro. In addition, the immune protection of ORF57 subunit vaccine was studied. The survival rate of CyHV-3 was 75% after 4 weeks of immunization, and the survival rate of ORF57 subunit vaccine group was significantly higher than that of PBS and PET-32a (+) groups. The results showed that the recombinant ORF57 subunit vaccine had strong immune protection, which provided an experimental basis for the clinical application of immunology and the development of CyHV-3 vaccine.