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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >High-level recombinant protein expression in transgenic plants by using a double-inducible viral vector
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High-level recombinant protein expression in transgenic plants by using a double-inducible viral vector

机译：双诱导病毒载体在转基因植物中高效表达重组蛋白

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摘要

We describe here a unique ethanol-inducible process for expression of recombinant proteins in transgenic plants. The process is based on inducible release of viral RNA replicons from stably integrated DNA proreplicons. A simple treatment with ethanol releases the replicon leading to RNA amplification and high-level protein production. To achieve tight control of replicon activation and spread in the uninduced state, the viral vector has been deconstructed, and its two components, the replicon and the cell-to-cell movement protein, have each been placed separately under the control of an inducible promoter. Transgenic Nicotiana benthamiana plants incorporating this double-inducible system demonstrate negligible background expression, high (over 0.5 × 10~4-fold) induction multiples, and high absolute levels of protein expression upon induction (up to 4.3 mg/g fresh biomass). The process can be easily scaled up, supports expression of practically important recombinant proteins, and thus can be directly used for industrial manufacturing.
机译：我们在这里描述了一种独特的乙醇诱导过程，用于在转基因植物中表达重组蛋白。该过程基于从稳定整合的DNA原复制子中诱导释放病毒RNA复制子。用乙醇简单处理即可释放复制子，从而导致RNA扩增和高水平蛋白质生产。为了实现对复制子激活和在未诱导状态下扩散的严格控制，病毒载体已被解构，其两个组件复制子和细胞间移动蛋白已分别置于诱导型启动子的控制下。结合了这种双重诱导系统的转基因烟草本生植物显示出可忽略的背景表达，高的诱导倍数（超过0.5×10〜4倍）和诱导后的蛋白质绝对表达水平高（最高为4.3 mg / g新鲜生物量）。该过程可以轻松扩展规模，支持表达实际上重要的重组蛋白，因此可以直接用于工业生产。

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