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一种用于在嵌合竹花叶病毒颗粒上表达表位的转基因植物细胞悬浮系统。

A transgenic plant cell-suspension system for expression of epitopes on chimeric Bamboo mosaic virus particles.

作者信息

Muthamilselvan Thangarasu, Lee Chin-Wei, Cho Yu-Hsin, Wu Feng-Chao, Hu Chung-Chi, Liang Yu-Chuan, Lin Na-Sheng, Hsu Yau-Heiu

机构信息

Graduate Institute of Biotechnology, National Chung Hsing University, Taichung, Taiwan.

Agricultural Biotechnology Research Center, Academia Sinica, Nankang, Taipei, Taiwan.

出版信息

Plant Biotechnol J. 2016 Jan;14(1):231-9. doi: 10.1111/pbi.12377. Epub 2015 Apr 16.

Abstract

We describe a novel strategy to produce vaccine antigens using a plant cell-suspension culture system in lieu of the conventional bacterial or animal cell-culture systems. We generated transgenic cell-suspension cultures from Nicotiana benthamiana leaves carrying wild-type or chimeric Bamboo mosaic virus (BaMV) expression constructs encoding the viral protein 1 (VP1) epitope of foot-and-mouth disease virus (FMDV). Antigens accumulated to high levels in BdT38 and BdT19 transgenic cell lines co-expressing silencing suppressor protein P38 or P19. BaMV chimeric virus particles (CVPs) were subsequently purified from the respective cell lines (1.5 and 2.1 mg CVPs/20 g fresh weight of suspended biomass, respectively), and the resulting CVPs displayed VP1 epitope on the surfaces. Guinea pigs vaccinated with purified CVPs produced humoral antibodies. This study represents an important advance in the large-scale production of immunopeptide vaccines in a cost-effective manner using a plant cell-suspension culture system.

摘要

我们描述了一种使用植物细胞悬浮培养系统来生产疫苗抗原的新策略,以替代传统的细菌或动物细胞培养系统。我们从携带编码口蹄疫病毒(FMDV)病毒蛋白1(VP1)表位的野生型或嵌合竹花叶病毒(BaMV)表达构建体的本氏烟草叶片中生成了转基因细胞悬浮培养物。在共表达沉默抑制蛋白P38或P19的BdT38和BdT19转基因细胞系中,抗原积累到高水平。随后从各自的细胞系中纯化BaMV嵌合病毒颗粒(CVP)(分别为1.5和2.1 mg CVP/20 g悬浮生物质鲜重),所得CVP在表面展示VP1表位。用纯化的CVP免疫的豚鼠产生了体液抗体。这项研究代表了使用植物细胞悬浮培养系统以经济有效的方式大规模生产免疫肽疫苗方面的一项重要进展。

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