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融合种子贮藏蛋白基因的 3'UTR 可导致种子中大量重组蛋白的积累。

Fusing the 3'UTR of seed storage protein genes leads to massive recombinant protein accumulation in seeds.

机构信息

Laboratory of Organelle Regulation, National Institute for Basic Biology, Okazaki, 444-8585, Japan.

Department of Viral Pathogenesis and Controls, National Center for Global Health and Medicine, Ichikawa, 272-8516, Japan.

出版信息

Sci Rep. 2023 Jul 27;13(1):12217. doi: 10.1038/s41598-023-39356-3.

Abstract

The demand for recombinant proteins is rising dramatically, and effective production systems are currently being developed. The production of recombinant proteins in plants is a promising approach due to its low cost and low risk of contamination of the proteins with endotoxins or infectious agents from the culture serum. Plant seeds primarily accumulate seed storage proteins (SSPs), which are transcribed and translated from a few genes; therefore, the mechanism underlying SSP accumulation has been studied to help devise ways to increase recombinant protein production. We found that the 3'UTR of SSP genes are essential for SSP accumulation and can be used in the production of recombinant proteins in Arabidopsis. Fusion of the 3'UTR of SSP genes to the 3' ends of DNA sequences encoding recombinant proteins enables massive accumulation of recombinant proteins with enzymatic activity in Arabidopsis seeds. This method is also applicable to the production of human Interferon Lambda-3 (IFN-lambda 3), a candidate biopharmaceutical compound against hepatitis C infection. Considering the low cost and ease of protein production in Arabidopsis, as well as the rapid growth of this plant, our method is useful for large-scale preparation of recombinant proteins for both academic research and biopharmaceutical production.

摘要

对重组蛋白的需求正在急剧增长,目前正在开发有效的生产系统。由于植物生产重组蛋白的成本低,而且蛋白质中内毒素或来自培养血清的传染性物质污染的风险低,因此它是一种很有前途的方法。植物种子主要积累种子贮藏蛋白(SSP),这些蛋白由少数基因转录和翻译而来;因此,人们研究了 SSP 积累的机制,以帮助设计增加重组蛋白产量的方法。我们发现 SSP 基因的 3'UTR 对于 SSP 的积累是必不可少的,可以用于在拟南芥中生产重组蛋白。将 SSP 基因的 3'UTR 与编码重组蛋白的 DNA 序列的 3'末端融合,能够使具有酶活性的重组蛋白在拟南芥种子中大量积累。该方法也适用于生产人干扰素 λ-3(IFN-lambda 3),这是一种针对丙型肝炎感染的生物制药候选化合物。考虑到在拟南芥中生产蛋白质的低成本和便利性,以及该植物的快速生长,我们的方法对于学术研究和生物制药生产中大规模制备重组蛋白都非常有用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a7cc/10374616/1bfd4017fcbf/41598_2023_39356_Fig1_HTML.jpg

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