Turpen T H
Biosource Technologies, Inc., Vacaville, CA 95688, USA.
Philos Trans R Soc Lond B Biol Sci. 1999 Mar 29;354(1383):665-73. doi: 10.1098/rstb.1999.0419.
There is a growing realization that a modern combination of molecular biology and agriculture will provide a photosynthetic basis for the biosynthesis of an increasing variety of complex and valuable molecules. This 'greening' of biotechnology may impact on the global environment in many beneficial ways, but will perhaps have its most significant impact on human health. In the past decade, the capacity to use plants as an expanded source of therapeutics has grown through the accelerated development of effective viral transfection vectors for gene transfer to cultivated crops. Recombinant vectors based on tobacco mosaic virus (TMV) and other members of the Tobamovirus genus are now used to transfect commercially meaningful quantities of plant biomass cultivated in enclosed greenhouses and multiacre fields. Viral RNA promoters are effectively manipulated for the synthesis of recombinant messenger RNAs in whole plants. Chimeric plant virus and virus-like particles are designed for peptide production and display from recombinant structural protein-gene fusions. Gene functions are assessed and modified by either virus-mediated expression or cytosolic inhibition of expression at the RNA level. Recombinant virus populations, propagated by inoculating plants with infectious RNA transcripts or recombinant virions, have proved to be genetically stable over product-manufacturing cycles. Large volumes of highly purified protein products isolated from transfected foliage conform reproducibly to the specifications required for well-characterized biologics. In some cases, they exceed the specific activities of molecules purified from alternative recombinant and native sources. The resulting products are then formulated according to the developing national regulatory guidelines appropriate for agriculture-based manufacturing. Each of these innovations was first realized by researchers using clones of tobamovirus genes and recombinant genomes. This progress is founded on the heritage of a century of fundamental TMV research.
人们越来越意识到，现代分子生物学与农业的结合将为越来越多复杂且有价值的分子的生物合成提供光合基础。生物技术的这种“绿色化”可能会以多种有益方式影响全球环境，但或许对人类健康的影响最为显著。在过去十年中，通过加速开发用于将基因转移到栽培作物的有效病毒转染载体，利用植物作为治疗药物扩展来源的能力得到了提升。基于烟草花叶病毒（TMV）和烟草花叶病毒属其他成员的重组载体现在被用于转染在封闭温室和多英亩田地中种植的具有商业意义数量的植物生物质。病毒RNA启动子被有效地用于在整株植物中合成重组信使RNA。嵌合植物病毒和病毒样颗粒被设计用于从重组结构蛋白 - 基因融合体中生产和展示肽。基因功能通过病毒介导的表达或RNA水平的胞质表达抑制来评估和修饰。通过用感染性RNA转录本或重组病毒粒子接种植物来繁殖的重组病毒群体，在产品制造周期中已被证明具有遗传稳定性。从转染的叶子中分离出的大量高度纯化的蛋白质产品可重复地符合特征明确的生物制品所需的规格。在某些情况下，它们超过了从替代重组和天然来源纯化的分子的比活性。然后根据适用于基于农业制造的国家监管指南制定所得产品。这些创新中的每一项最初都是由使用烟草花叶病毒基因克隆和重组基因组的研究人员实现的。这一进展建立在一个世纪的烟草花叶病毒基础研究的遗产之上。
