Gyeongik Ahn, Joon-Yung Cha, Jeong Won Lee, Gyeongran Park, Gyeong-Im Shin, Shi-Jian Song, Gyeongryul Ryu, Inhwan Hwang, Min Gab Kim, Woe-Yeon Kim
{"title":"利用本菌烟制备针对石鲷虹膜病毒的细菌样颗粒疫苗。","authors":"Gyeongik Ahn, Joon-Yung Cha, Jeong Won Lee, Gyeongran Park, Gyeong-Im Shin, Shi-Jian Song, Gyeongryul Ryu, Inhwan Hwang, Min Gab Kim, Woe-Yeon Kim","doi":"10.1007/s12374-021-09328-z","DOIUrl":null,"url":null,"abstract":"<p><p>Viral diseases are extremely widespread infections that change constantly through mutations. To produce vaccines against viral diseases, transient expression systems are employed, and <i>Nicotiana benthamiana</i> (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 <i>Lactococcus lactis</i>. The protein was abundantly produced in <i>N. benthamiana</i> in its <i>N</i>-glycosylated form. Total soluble proteins isolated from infiltrated <i>N. benthamiana</i> 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 OD<sub>600</sub> value of 0.4 trichloroacetic acid (TCA)-treated <i>L. lactis</i> 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 <i>L. lactis</i>. In summary, we developed a plant-based recombinant vaccine production system combined with surface display on <i>L. lactis</i>.</p>","PeriodicalId":16762,"journal":{"name":"Journal of Plant Biology","volume":"65 1","pages":"21-28"},"PeriodicalIF":2.2000,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8477727/pdf/","citationCount":"1","resultStr":"{\"title\":\"Production of a Bacteria-like Particle Vaccine Targeting Rock Bream (<i>Oplegnathus fasciatus</i>) Iridovirus Using <i>Nicotiana benthamiana</i>.\",\"authors\":\"Gyeongik Ahn, Joon-Yung Cha, Jeong Won Lee, Gyeongran Park, Gyeong-Im Shin, Shi-Jian Song, Gyeongryul Ryu, Inhwan Hwang, Min Gab Kim, Woe-Yeon Kim\",\"doi\":\"10.1007/s12374-021-09328-z\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Viral diseases are extremely widespread infections that change constantly through mutations. To produce vaccines against viral diseases, transient expression systems are employed, and <i>Nicotiana benthamiana</i> (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 <i>Lactococcus lactis</i>. The protein was abundantly produced in <i>N. benthamiana</i> in its <i>N</i>-glycosylated form. Total soluble proteins isolated from infiltrated <i>N. benthamiana</i> 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 OD<sub>600</sub> value of 0.4 trichloroacetic acid (TCA)-treated <i>L. lactis</i> 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 <i>L. lactis</i>. In summary, we developed a plant-based recombinant vaccine production system combined with surface display on <i>L. lactis</i>.</p>\",\"PeriodicalId\":16762,\"journal\":{\"name\":\"Journal of Plant Biology\",\"volume\":\"65 1\",\"pages\":\"21-28\"},\"PeriodicalIF\":2.2000,\"publicationDate\":\"2022-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8477727/pdf/\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Plant Biology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1007/s12374-021-09328-z\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2021/9/28 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q2\",\"JCRName\":\"PLANT SCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Plant Biology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1007/s12374-021-09328-z","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2021/9/28 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"PLANT SCIENCES","Score":null,"Total":0}
Production of a Bacteria-like Particle Vaccine Targeting Rock Bream (Oplegnathus fasciatus) Iridovirus Using Nicotiana benthamiana.
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.
期刊介绍:
Journal of Plant Biology, an official publication of the Botanical Society of Korea, is an international journal devoted to basic researches in biochemistry, cellular biology, development, ecology, genetics, molecular biology, physiology, and systematics of plants.
The Journal publishes the following categories of paper:
Original articles -- For publication in Journal of Plant Biology the manuscript must provide a significant new contribution to our understanding of plants. All areas of plant biology are welcome. No limit on the length, but a concise presentation is encouraged.
Reviews -- Invited by the EiC.
Brief Communications -- Concise but independent report representing significant contribution to plant science.
The Botanical Society of Korea was founded on November 30, 1957 to promote studies, disseminate and exchange information on the field of plant biology. The first issue of The Korean Journal of Botany, the official publication of the society, was published on April 1, 1958. It was published twice a year, but quarterly from 5th volume in 1962. In 1994, it was renamed to Journal of Plant Biology and published in English since 1996. The journal entered its 50th year of publication in 2007.