ABSTRACT Viral hemorrhagic septicemia (VHS) is an important infectious disease in fish worldwide caused by viral hemorrhagic septicemia virus (VHSV). VHSV is the causative agent of serious systemic diseases in fish, affecting a number of teleost fish species. In this study, VHSV glycoprotein (G), including its epitope, as a subunit vaccine candidate, was expressed in tobacco plant (Nicotiana tabacum). The recombinant gene, VHSVG, was fused to the immunoglobulin Fc fragment and extended with the endoplasmic reticulum (ER) retention signal (KDEL) to generate VHSVG-FcK. The recombinant expression vector for VHSVG-FcK was transferred into Agrobacterium tumefaciens (LBA4404), and plant transformation was conducted N. tabacum. Polymerase chain reaction (PCR) was performed to confirm gene insertion and VHSVG-FcK protein expression was confirmed by immunoblot analysis. VHSVG-FcK protein was successfully purified from tobacco plant leaves. Furthermore, ELISA analysis showed that mice serum immunized with the plant-derived VHSVG-FcK (VHSVGP-FcK) had a high absorbance against VHSVG-FcK, indicating that the plant-derived recombinant subunit vaccine protein VHSVG-FcK can induce immune response. Taken together, this recombinant vaccine protein can be expressed in plant expression systems and can be appropriately assembled to be functional in immunogenicity.

中文翻译：

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病毒性出血性败血症病毒（VHSV）异源重组抗原蛋白在小鼠体内的免疫反应

【摘要】：病毒性出血性败血症(VHS)是由病毒性出血性败血症病毒(VHSV)引起的全球鱼类重要传染病。VHSV 是鱼类严重全身性疾病的病原体，影响许多硬骨鱼类。在这项研究中，VHSV 糖蛋白 (G)，包括其表位，作为亚单位疫苗候选者，在烟草植物 (Nicotiana tabacum) 中表达。重组基因 VHSVG 与免疫球蛋白 Fc 片段融合，并用内质网 (ER) 保留信号 (KDEL) 延伸以产生 VHSVG-FcK。将VHSVG-FcK重组表达载体转入根癌农杆菌(LBA4404)，进行植物转化。进行聚合酶链反应 (PCR) 以确认基因插入，并通过免疫印迹分析确认 VHSVG-FcK 蛋白表达。VHSVG-FcK 蛋白从烟草植物叶片中成功纯化。此外，ELISA分析表明，用植物源性VHSVG-FcK（VHSVGP-FcK）免疫的小鼠血清对VHSVG-FcK具有高吸光度，表明植物源性重组亚单位疫苗蛋白VHSVG-FcK可以诱导免疫反应。总之，这种重组疫苗蛋白可以在植物表达系统中表达，并且可以适当组装以具有免疫原性功能。ELISA分析表明，用植物源性VHSVG-FcK（VHSVGP-FcK）免疫的小鼠血清对VHSVG-FcK具有高吸光度，表明植物源性重组亚单位疫苗蛋白VHSVG-FcK可诱导免疫反应。总之，这种重组疫苗蛋白可以在植物表达系统中表达，并且可以适当组装以具有免疫原性功能。ELISA分析表明，用植物源性VHSVG-FcK（VHSVGP-FcK）免疫的小鼠血清对VHSVG-FcK具有高吸光度，表明植物源性重组亚单位疫苗蛋白VHSVG-FcK可诱导免疫反应。总之，这种重组疫苗蛋白可以在植物表达系统中表达，并且可以适当组装以具有免疫原性功能。