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KMID : 0368420220650010021
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Journal of Plant Biology
2022 Volume.65 No. 1 p.21 ~ p.28
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Production of a Bacteria-like Particle Vaccine Targeting Rock Bream (Oplegnathus fasciatus) Iridovirus Using Nicotiana benthamiana
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Ahn Gyeong-Ik
Cha Joon-Yung
Lee Jeong-Won
Park Gyeong-Ran
Shin Gyeong-Im
Song Shi-Jian
Ryu Gyeong-Ryul
Hwang In-Hwan
Kim Min-Gab
Kim Woe-Yeon
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Abstract
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Viral diseases are extremely widespread infections that change constantly through mutations. To produce vaccines against viral diseases, transient expression systems are employed, and Nicotiana benthamiana (tobacco) plants are a rapidly expanding platform. In this study, we developed a recombinant protein vaccine targeting the major capsid protein (MCP) of iridovirus fused with the lysine motif (LysM) and coiled-coil domain of coronin 1 (ccCor1) for surface display using Lactococcus lactis. The protein was abundantly produced in N. benthamiana in its N-glycosylated form. Total soluble proteins isolated from infiltrated N. benthamiana leaves were treated sequentially with increasing ammonium sulfate solution, and recombinant MCP mainly precipitated at 40?60%. Additionally, affinity chromatography using Ni?NTA resin was applied for further purification. Native structure analysis using size exclusion chromatography showed that recombinant MCP existed in a large oligomeric form. A minimum OD600 value of 0.4 trichloroacetic acid (TCA)-treated L. lactis was required for efficient recombinant MCP display. Immunogenicity of recombinant MCP was assessed in a mouse model through enzyme-linked immunosorbent assay (ELISA) with serum-injected recombinant MCP-displaying L. lactis. In summary, we developed a plant-based recombinant vaccine production system combined with surface display on L. lactis.
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KEYWORD
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Bacteria-like particle, Iridovirus, Major capsid protein, Nicotiana benthamiana, Plant-based vaccine
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