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

Preparation and immunoprotection of ORF57 subunit vaccine of Cyprinid herpesvirus 3

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。

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.