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.