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Open AccessArticle

Production of Human IFNγ Protein in Nicotiana benthamiana Plant through an Enhanced Expression System Based on Bamboo mosaic Virus

1
Ph.D Program in Microbial Genomic, National Chung Hsing University and Academia Sinica, Taichung 40227, Taiwan
2
Graduate Institute of Biotechnology, Advanced Plant Biotechnology Center, National Chung Hsing University, Taichung 40227, Taiwan
3
Institute of Plant and Microbial Biology, Academia Sinica, Taipei 11529, Taiwan
*
Author to whom correspondence should be addressed.
Viruses 2019, 11(6), 509; https://doi.org/10.3390/v11060509
Received: 8 May 2019 / Revised: 30 May 2019 / Accepted: 1 June 2019 / Published: 3 June 2019
(This article belongs to the Section Viruses of Plants, Fungi and Protozoa)
Plant-based systems are safe alternatives to the current platforms for the production of biologically active therapeutic proteins. However, plant-based expression systems face certain major challenges, including the relatively low productivity and the generation of target proteins in biologically active forms. The use of plant virus-based expression systems has been shown to enhance yields, but further improvement is still required to lower the production cost. In this study, various strategies were employed to increase the yields of an important therapeutic protein, human interferon gamma (IFNγ), in Nicotiana benthamiana through modifications of expression vectors based on potexviruses. Among these, the vector based on a coat protein (CP)-deficient Bamboo mosaic virus (BaMV), pKB△CHis, was shown to exhibit the highest expression level for the unmodified IFNγ. Truncation of the N-terminal signal peptide of IFN (designated mIFNγ) resulted in a nearly seven-fold increase in yield. Co-expression of a silencing suppressor protein by replacing the coding sequence of BaMV movement protein with that of P19 led to a 40% increase in mIFNγ accumulation. The fusion of endoplasmic reticulum (ER) retention signal with mIFNγ significantly enhanced the accumulation ratio of biologically active dimeric mIFNγ to 87% relative to the non-active monomeric form. The construct pKB19mIFNγER, employing the combination of all the above enhancement strategies, gave the highest level of protein accumulation, up to 119 ± 0.8 μg/g fresh weight, accounting for 2.5% of total soluble protein (TSP) content. These findings advocate the application of the modified BaMV-based vector as a platform for high-level expression of therapeutic protein in N. benthamiana. View Full-Text
Keywords: potexvirus; bamboo mosaic virus; interferon gamma; therapeutic protein; ER retention potexvirus; bamboo mosaic virus; interferon gamma; therapeutic protein; ER retention
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MDPI and ACS Style

Jiang, M.-C.; Hu, C.-C.; Lin, N.-S.; Hsu, Y.-H. Production of Human IFNγ Protein in Nicotiana benthamiana Plant through an Enhanced Expression System Based on Bamboo mosaic Virus. Viruses 2019, 11, 509.

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