A method for rapid production of heteromultimeric protein complexes in plants: assembly of protective bluetongue virus-like particles
A method for rapid production of heteromultimeric protein complexes in plants: assembly of protective bluetongue virus-like particles
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DOI：
10.1111/pbi.12076
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发表时间：
2013-09-01
影响因子：
13.8
通讯作者：
Lomonossoff, George P.
中科院分区：
文献类型：
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作者：
Thuenemann, Eva C.;Meyers, Ann E.;Lomonossoff, George P.
基于非复制型病毒载体的植物表达系统可用于在同一细胞内同时表达多个基因。因此，它们具有生产异多聚体蛋白质复合物的巨大潜力。这项工作描述了蓝舌病病毒样颗粒（VLP）的有效植物生产和组装，需要同时表达不同数量的四种不同蛋白质。这种颗粒有可能作为抗蓝舌病病毒（BTV）的安全有效的疫苗，这种病毒会导致反刍动物的高死亡率，从而对牲畜贸易产生严重影响。在这里，VLP生产和组装在烟草本塞姆亚纳使用豇豆花叶病毒为基础的HyperTrans（CPMV-HT）和相关的pEAQ植物瞬时表达载体系统显示，在绵羊中引发强烈的抗体反应。此外，它们提供了针对南非BTV-8田间分离株攻击的保护性免疫。结果表明，瞬时表达可用于在几天内在植物中产生免疫相关的复杂异源多聚体结构。该结果具有超出兽医疫苗领域的影响，并可应用于生产供人类使用的VLP或共表达多种酶以操纵代谢途径。
Plant expression systems based on nonreplicating virus-based vectors can be used for the simultaneous expression of multiple genes within the same cell. They therefore have great potential for the production of heteromultimeric protein complexes. This work describes the efficient plant-based production and assembly of Bluetongue virus-like particles (VLPs), requiring the simultaneous expression of four distinct proteins in varying amounts. Such particles have the potential to serve as a safe and effective vaccine against Bluetongue virus (BTV), which causes high mortality rates in ruminants and thus has a severe effect on the livestock trade. Here, VLPs produced and assembled in Nicotiana benthamiana using the cowpea mosaic virus-based HyperTrans (CPMV-HT) and associated pEAQ plant transient expression vector system were shown to elicit a strong antibody response in sheep. Furthermore, they provided protective immunity against a challenge with a South African BTV-8 field isolate. The results show that transient expression can be used to produce immunologically relevant complex heteromultimeric structures in plants in a matter of days. The results have implications beyond the realm of veterinary vaccines and could be applied to the production of VLPs for human use or the coexpression of multiple enzymes for the manipulation of metabolic pathways.