In the past decade, interest in the production of recombinant pharmaceutical proteins in plants has tremendously progressed because plants do not harbor mammalian viruses, are economically competitive, easily scalable, and capable of carrying out complex post-translational modifications required for recombinant pharmaceutical proteins. Mucuna bracteata is an essential perennial cover crop species widely planted as an underground cover in oil palm and rubber plantations. As a legume, they have high biomass, thrive in its habitat, and can fix nitrogen. Thus, M. bracteata is a cost-efficient crop that shows ideal characteristics as a platform for mass production of recombinant protein. In this study, we established a new platform for the transient production of a recombinant protein in M. bracteata via vacuum-assisted agro-infiltration. Five-week-old M. bracteata plants were vacuum infiltrated with Agrobacterium tumefaciens harboring a plasmid that encodes for an anti-toxoplasma immunoglobulin (IgG) under different parameters, including trifoliate leaf positional effects, days to harvest post-infiltration, and the Agrobacterium strain used. Our results showed that vacuum infiltration of M. bracteata plant with A. tumefaciens strain GV3101 produced the highest concentration of heterologous protein in its bottom trifoliate leaf at 2 days post-infiltration. The purified anti-toxoplasma IgG was then analyzed using Western blot and ELISA. It was demonstrated that, while structural heterogeneity existed in the purified anti-toxoplasma IgG from M. bracteata, its transient expression level was two-fold higher than the model platform, Nicotiana benthamiana. This study has laid the foundation towards establishing M. bracteata as a potential platform for the production of recombinant pharmaceutical protein.

中文翻译：

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用于生产药物蛋白的新型植物表达系统。

在过去的十年中，由于植物不携带哺乳动物病毒，经济上具有竞争力，易于扩展并且能够进行重组药物蛋白所需的复杂翻译后修饰，因此在植物中生产重组药物蛋白的兴趣得到了极大的发展。Mucuna bracteata是一种重要的多年生覆盖作物，广泛用作油棕和橡胶种植园的地下覆盖。作为一种豆科植物，它们具有较高的生物量，在其栖息地中壮成长，并且可以固氮。因此，Bracteata是一种经济高效的作物，具有理想的特性，可作为大规模生产重组蛋白的平台。在这项研究中，我们建立了一个新的平台，用于通过真空辅助农业浸润法在短小分支杆菌中瞬时生产重组蛋白。将五周龄的Bras bracteata植物用根癌土壤杆菌真空浸润，该土壤杆菌带有一个质粒，该质粒在不同参数下编码抗弓形虫免疫球蛋白（IgG），包括三叶叶叶片的位置效应，浸润后收获的天数和土壤杆菌菌株用过的。我们的结果表明，在浸入后2天，根癌农杆菌GV3101对Bracteata植物的真空浸渗产生了最高浓度的异源蛋白质。然后使用Western blot和ELISA分析纯化的抗弓形虫IgG。结果表明，虽然纯化的弓形虫抗弓形虫IgG存在结构异质性，但其瞬时表达水平比模型平台Nicotiana benthamiana高两倍。