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Enhanced expression of dengue virus EDIII-based tetravalent antigen protein using transgenic rice callus

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Abstract

Mosquito-borne tropical dengue disease is a major global epidemic affecting hundreds of millions of people. As a precaution, it would be ideal to develop a highly efficient tetravalent vaccine that protects against all serotypes of dengue viruses. In this study, envelope protein domain III (EDIII) of dengue virus was employed as a target antigen. A single-chain tetravalent type of EDIII (tEDIII) was generated by utilizing a linker peptide and transformed into rice (Oryza sativa). Integration and mRNA transcripts of the tEDIII gene were confirmed by genomic DNA polymerase chain reaction (PCR) and Northern blot analyses, respectively. The expression of wild-type tEDIII (wtEDIII) protein was confirmed by Western blot analysis and was determined to improve the synthesis of the tEDIII (stEDIII) construct based on codon optimization and ER targeting. The yield of stEDIII protein was sevenfold higher than secreted wtEDIII protein, reaching a maximum of 357 µg per gram dry weight. These results suggest that a simple tetravalent EDIII dengue antigen can be produced in rice, raising the possibility that edible plant cells can be vaccinated by mucosal application for protection against dengue infection.

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Acknowledgements

This study was supported by NRF-2014K1B1A1073861 through the National Research Foundation (NRF) funded by the Korean Ministry of Science, ICT & Future Planning, and by the Ministry of Agriculture, Food and Rural Affairs (714001-07), Republic of Korea.

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Authors and Affiliations

  1. Department of Molecular Biology, Chonbuk National University, Jeonju, Republic of Korea

    So-Chon Han, Nguyen-Xuan Huy & Mi-Young Kim

  2. Biology Department, University of Education, Hue University, 34 Le Loi, Hue, Vietnam

    Nguyen-Xuan Huy

  3. Jeonbuk Institute for Food-Bioindustry, Jeonju, Republic of Korea

    So-Chon Han

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  1. So-Chon Han
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  2. Nguyen-Xuan Huy
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M-YK conceived and designed the overall study. S-CH and N-XH performed the majority of experiments. S-CH and M-YK wrote the manuscript.

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Correspondence to Mi-Young Kim.

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Han, SC., Huy, NX. & Kim, MY. Enhanced expression of dengue virus EDIII-based tetravalent antigen protein using transgenic rice callus. Plant Biotechnol Rep 12, 207–215 (2018). https://doi.org/10.1007/s11816-018-0487-6

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