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鸡胚生物反应器中重组蛋白生产表达系统的评价。

Evaluation of expression systems for recombinant protein production in chicken egg bioreactors.

机构信息

Laboratory of Immunobiology, Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima, Hiroshima, Japan.

Genome Editing Innovation Center, Hiroshima University, Higashi-Hiroshima, Hiroshima, Japan.

出版信息

Biotechnol J. 2024 Jan;19(1):e2300316. doi: 10.1002/biot.202300316. Epub 2023 Oct 27.

Abstract

Chicken eggs have gained attention as excellent bioreactors because of their genetic modifications. However, the development of chicken egg bioreactors requires a long time from the construction of the production system to the evaluation of the products. Therefore, in this study, a chicken cell line producing ovalbumin (OVA) was established and constructed a system for the rapid evaluation of the production system. First, the EF1α promoter was knocked in upstream of the OVA locus in chicken DF-1 cells for continuous OVA expression. Furthermore, an ideal position at the OVA locus for the insertion of useful protein genes to maximize recombinant protein yield was analyzed and identified. The knocking in the EF1α promoter upstream of exon1 yielded the maximum production of OVA protein was achieved. In addition, Linking a recombinant hFGF2 cDNA to the 5' side of the OVA was found to increase production efficiency. Therefore, an OVA-expressing cell line and an evaluation system for proteins in chicken egg bioreactors was established. The findings may improve the efficiency of chicken expression systems and expand their applications in protein production.

摘要

鸡蛋作为优秀的生物反应器,因其遗传修饰而备受关注。然而,从生产系统的构建到产品的评估,鸡卵生物反应器的开发需要很长时间。因此,在本研究中,建立了一种生产卵清白蛋白(OVA)的鸡细胞系,并构建了一种快速评估生产系统的系统。首先,将 EF1α 启动子敲入鸡 DF-1 细胞的 OVA 基因座上游,用于持续表达 OVA。此外,分析并确定了在 OVA 基因座中插入有用蛋白基因的理想位置,以最大限度地提高重组蛋白的产量。在 exon1 的上游敲入 EF1α 启动子可获得最大的 OVA 蛋白产量。此外,将重组 hFGF2 cDNA 连接到 OVA 的 5' 端可提高生产效率。因此,建立了一种表达 OVA 的细胞系和鸡卵生物反应器中蛋白质的评估系统。这些发现可能会提高鸡表达系统的效率,并扩大其在蛋白质生产中的应用。

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