Transient protein expression in three Pisum sativum (green pea) varieties
Transient protein expression in three Pisum sativum (green pea) varieties
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DOI：
10.1002/biot.200800256
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发表时间：
2009-02-01
影响因子：
4.7
通讯作者：
Mett, Vadim
中科院分区：
文献类型：
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作者：
Green, Brian J.;Fujiki, Masaaki;Mett, Vadim
植物中蛋白质的瞬时表达和通过永久转化系的表达已被许多研究组证实。瞬时植物表达系统，由于高表达水平和生产速度，与永久转化体相比，显示出更大的生物制药生产前景。基于烟草花叶病毒（TMV）的表达载体是最常用的，并且所使用的主要植物本塞姆氏烟草（Nicotianabenthamiana）已经证明能够以适于纯化的水平表达广泛的蛋白质。N. benthamiana的使用有两个限制：一个是生长相对缓慢，另一个是生物量低。为了解决这些局限性，我们筛选了一些豆科植物的瞬时蛋白表达。使用苜蓿花叶病毒（AMV）和黄瓜花叶病毒（CMV）载体，通过农杆菌，我们能够确定三个豌豆品种，表现出蛋白质表达瞬时。在绿色豌豆品种斑点豌豆中达到420 +/-26.24 mg CFP/kgFW的表达水平。我们还能够表达三种治疗性蛋白质，这表明该系统在生物制药生产中的前景。
The expression of proteins in plants both transiently and via permanently transformed lines has been demonstrated by a number of groups. Transient plant expression systems, due to high expression levels and speed of production, show greater promise for the manufacturing of biopharmaceuticals when compared to permanent transformants. Expression vectors based on a tobacco mosaic virus (TMV) are the most commonly utilized and the primary plant used, Nicotiana benthamiana, has demonstrated the ability to express a wide range of proteins at levels amenable to purification. N. benthamiana has two limitations for its use; one is its relatively slow growth, and the other is its low biomass. To address these limitations we screened a number of legumes for transient protein expression. Using the alfalfa mosaic virus (AMV) and the cucumber mosaic virus (CMV) vectors, delivered via Agrobacterium, we were able to identify three Pisum sativum varieties that demonstrated protein expression transiently. Expression levels of 420 +/- 26.24 mg CFP/kgFW in the green pea variety speckled pea were achieved. We were also able to express three therapeutic proteins indicating promise for this system in the production of biopharmaceuticals.