Matsuo Kouki, Hong Jin-Sung, Tabayashi Noriko, Ito Akira, Masuta Chikara, Matsumura Takeshi
National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Sapporo, Japan.
Planta. 2007 Jan;225(2):277-86. doi: 10.1007/s00425-006-0346-5. Epub 2006 Jul 5.
We have developed Cucumber mosaic virus (CMV) as a plant virus vector especially for production of pharmaceutical proteins. The CMV vector is a vector modifiable for different host plants and does not require further engineering steps. CMV contains three genomic RNA molecules (RNAs 1-3) necessary for infectivity. With this system, instead of creating different vector constructs for each plant we use, we take advantage of the formation of pseudrecombinants between two CMV isolates by simply reassembling a vector construct (RNA 2 base) and an RNA molecule containing the host determinant (mostly RNA 3). In this study, the gene for acidic fibroblast growth factor (aFGF), one of the human cytokines, was cloned under the control of the subgenomic promoter for RNA 4A of the CMV-based vector, C2-H1. Infected Nicotiana benthamiana plants produced aFGF at levels up to 5-8% of the total soluble protein. The tobacco-produced aFGF was purified, and its biological activity was confirmed. Using this system, which provides a versatile and viable strategy for the production of therapeutic proteins in plants, we also demonstrated a high level of aFGF in Glycine max (soybean) and Arabidopsis thaliana.
我们已开发出黄瓜花叶病毒（CMV）作为一种植物病毒载体，专门用于生产药用蛋白质。CMV载体是一种可针对不同宿主植物进行改造的载体，无需进一步的工程步骤。CMV含有感染性所必需的三个基因组RNA分子（RNA 1 - 3）。利用该系统，我们不是为每种使用的植物构建不同的载体构建体，而是通过简单地重新组装一个载体构建体（RNA 2碱基）和一个含有宿主决定簇的RNA分子（大多为RNA 3），利用两种CMV分离株之间形成假重组体的特性。在本研究中，人类细胞因子之一的酸性成纤维细胞生长因子（aFGF）基因在基于CMV的载体C2 - H1的RNA 4A亚基因组启动子的控制下被克隆。被感染的本氏烟草植物产生的aFGF水平高达总可溶性蛋白的5 - 8%。对烟草产生的aFGF进行了纯化，并证实了其生物活性。利用该系统为在植物中生产治疗性蛋白质提供了一种通用且可行的策略，我们还在大豆和拟南芥中展示了高水平表达的aFGF。
