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Molecular Farming, an Effective System for the Production of Immunogenic Crimean-Congo Hemorrhagic Fever Virus Glycoprotein | ||
| Progress in Biological Sciences | ||
| مقاله 2، دوره 2، شماره 1، خرداد 2012، صفحه 12-29 اصل مقاله (1.15 M) | ||
| شناسه دیجیتال (DOI): 10.22059/pbs.2012.24821 | ||
| نویسندگان | ||
| Seyed Mojtaba Ghiasi1؛ Ali-Hatef Salmanian1؛ Ali Sharafi1؛ Ruhollah Kazemi1؛ Mahya Jafari1؛ Sadegh Chinikar2؛ Sedighe Zakeri2 | ||
| 1National Institute of Genetic Engineering and Biotechnology | ||
| 2Pasteur Institute of Iran, Tehran | ||
| چکیده | ||
| 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. | ||
| کلیدواژه‌ها | ||
| CCHFV؛ Molecular farming؛ Transgenic plant | ||
| عنوان مقاله [English] | ||
| Molecular Farming, an Effective System for the Production of Immunogenic Crimean-Congo Hemorrhagic Fever Virus Glycoprotein | ||
| نویسندگان [English] | ||
| Seyed Mojtaba Ghiasi1؛ Ali-Hatef Salmanian1؛ Ali Sharafi1؛ Ruhollah Kazemi1؛ Mahya Jafari1؛ Sadegh Chinikar2؛ Sedighe Zakeri2 | ||
| چکیده [English] | ||
| 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. | ||
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