High expression of consensus dengue virus envelope glycoprotein domain III using a viral expression system in tobacco
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Although plant-based vaccines have many advantages, their use is limited by low expression of antigen genes in transgenic plants, which results in low immune responses and immune tolerance. To overcome this problem, Nicotiana benthamiana was used to produce the consensus domain III of dengue virus envelope glycoprotein (E) via agroinfiltration with a plant virus-based expression system. The binding of E glycoprotein to a receptor is important for dengue virus entry into host cells and results in human disease. Consensus domain III of dengue virus E glycoprotein (cEDIII) is immunogenic and can elicit neutralizing antibodies against all four serotypes of dengue virus. A DNA fragment encoding cEDIII and M cell-targeting ligand fused to cEDIII (cEDIII-Co1) were constructed in a viral vector and introduced into tobacco plant cells by Agrobacterium-mediated infiltration. The cEDIII and cEDIII-Co1 fusion proteins were detected in protein extracts from agroinfiltrated leaves by Western blot analysis. The plant-produced cEDIII and cEDIII-Co1 fusion proteins composed 5.2 and 4.8 mg/g of dry weight of leaf tissues, respectively. These results suggest that the high expression of dengue virus cEDIII in plants with a plant virus-based expression system can overcome the low expression level to improve the feasibility of plant-based vaccines.
KeywordsDengue virus Plant-based vaccine Plant viral expression systems Consensus domain III
This paper was supported by a research fund from Chonbuk National University in 2014 and by the National Research Foundation (NRF-2014K1B1A1073861) funded by Korean Ministry of Science, ICT and Future Planning.
- Maclean J, Koekemoer M, Olivier AJ, Stewart D, Hitzeroth II, Rademacher T, Fischer R, Williamson AL, Rybicki EP (2007) Optimization of human papillomavirus type 16 (HPV-16) L1 expression in plants: comparison of the suitability of different HPV-16 L1 gene varients and different cell-compartment localization. J Gen Virol 88:1460–1469PubMedCrossRefGoogle Scholar