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.
Plant molecular biofarming has been increasingly studied in recent decades. Many kinds of recombinant immunotherapeutic proteins have been produced in transgenic tobacco plants. However, tobacco mosaic virus (TMV) can damage tobacco plants and cause pathogenic symptoms, which affects plant biomass production. Cigarette sap solution has been used to infect TMV during tobacco cultivation. In this study, we obtained TMV-infected transgenic plants expressing a prostatespecific antigen (PSA) fused to an IgG Fc with KDEL ER retention motif (PSA-IgG FcK). The typical TMV-associated symptoms appeared and increased on plant leaves 5 days after infection. mRNA expression levels and the presence of TMV were confirmed by RT-PCR and transmission electron microscopy analyses. The expression level and purity of the target protein were not significantly different between noninfected and infected transgenic plants. TMV was not found in the purified protein samples from infected plants. Our study showed that TMV pathogen-infected plant biomass can be harvested and processed to obtain therapeutic vaccine proteins.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.