Higher accumulation of F1-V fusion recombinant protein in plants after induction of protein body formation
Higher accumulation of F1-V fusion recombinant protein in plants after induction of protein body formation
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DOI：
10.1007/s11103-009-9552-4
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发表时间：
2010-01-01
影响因子：
5.1
通讯作者：
Cardineau, Guy A.
中科院分区：
文献类型：
--
作者：
Alvarez, M. Lucrecia;Topal, Emel;Cardineau, Guy A.
提高外源蛋白的积累对于提高植物性生产系统的商业成功至关重要，因为产品产量对过程经济学有重大影响。谷物进化成以紧密组织的聚集体储存大量蛋白质。在玉米中，γ - zein是由粗内质网（ER）合成并储存在称为蛋白体（PB）的特殊细胞器中的主要储存蛋白。Zera(A (R)) （γ -zein er - accumulation domain）是γ -zein的n端富含脯氨酸的结构域，足以诱导PB形成的组装。Zera（A (R)）结构域与感兴趣的蛋白质融合导致密集的pb样er衍生细胞器的组装，其中含有高浓度的重组蛋白。我们的主要目标是增加重组蛋白在植物中的积累，以提高口服植物制造疫苗的效率。众所周知，口服疫苗接种所需的剂量比亲本配方高得多。作为开发植物鼠疫疫苗项目的一部分，我们表达了我们的模型抗原，鼠疫耶尔森菌F1-V抗原融合蛋白，有和没有融合的Zera（a(R)）结构域。我们证明，Zera(A (R))-F1-V蛋白在三种不同的宿主植物系统中表达时，积累量至少比F1-V单独表达高3倍：Ncotiana benthamiana， Medicago sativa（苜蓿）和Nicotiana tabacum NT1细胞。我们证实了Zera（A (R)）技术在非种子组织中诱导蛋白体形成的可行性。Zera（A (R)）的表达和积累不影响植株的发育和生长。这些结果证实了Zera（A (R)）技术在大幅增加植物增值蛋白积累方面的潜力。
Improving foreign protein accumulation is crucial for enhancing the commercial success of plant-based production systems since product yields have a major influence on process economics. Cereal grain evolved to store large amounts of proteins in tightly organized aggregates. In maize, gamma-Zein is the major storage protein synthesized by the rough endoplasmic reticulum (ER) and stored in specialized organelles called protein bodies (PB). Zera(A (R)) (gamma-Zein ER-accumulating domain) is the N-terminal proline-rich domain of gamma-zein that is sufficient to induce the assembly of PB formation. Fusion of the Zera(A (R)) domain to proteins of interest results in assembly of dense PB-like, ER-derived organelles, containing high concentration of recombinant protein. Our main goal was to increase recombinant protein accumulation in plants in order to enhance the efficiency of orally-delivered plant-made vaccines. It is well known that oral vaccination requires substantially higher doses than parental formulations. As a part of a project to develop a plant-made plague vaccine, we expressed our model antigen, the Yersinia pestis F1-V antigen fusion protein, with and without a fused Zera(A (R)) domain. We demonstrated that Zera(A (R))-F1-V protein accumulation was at least 3x higher than F1-V alone when expressed in three different host plant systems: Ncotiana benthamiana, Medicago sativa (alfalfa) and Nicotiana tabacum NT1 cells. We confirmed the feasibility of using Zera(A (R)) technology to induce protein body formation in non-seed tissues. Zera(A (R)) expression and accumulation did not affect plant development and growth. These results confirmed the potential exploitation of Zera(A (R)) technology to substantially increase the accumulation of value-added proteins in plants.