molecular farming, an effective system for the production of immunogenic crimean-congo hemorrhagic fever virus glycoprotein

the main aim of this study was to obtain the crimean-congo hemorrhagic fever virus (cchfv) glycoprotein, through either stable transgenic plants or using a transient expression system, and determine the yield, quality and finally the immunogenicity of the plant-made cchfv glycoprotein in a mouse model. we designed and synthesized a codon-optimized g1/g2 gene from the g1 and g2 parts of the cchfv glycoprotein by bioinformatic analysis. the synthetic construct was cloned into a plant expression vector and tobacco plants were both transiently and stably transformed. the transgenic plantlets or tobacco-derived hairy roots confirmed by pcr and southern blot analyses. the intact 98 kda g1/g2 glycoprotein was produced by a transient expression system at as much as 3.3 mg/kg fresh weight. the recombinant g1/g2 protein was analyzed in stable lines by g1/g2 elisa and western blot. the yield in the transgenic hairy root line was significantly higher than that in transgenic tobacco lines. finally, the immunogenicity of the plant-made g1/g2 glycoprotein was evaluated by its subcutaneous administration in mice when compared with positive and negative controls. this plant-purified g1/g2 protein produced a high titer of anti-cchfv glycoprotein igg antibodies in mice.