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

IF 3.2 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS
Ryota Kajihara, Ryo Ezaki, Tenkai Watanabe, Kennosuke Ichikawa, Mei Matsuzaki, Hiroyuki Horiuchi
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引用次数: 0

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.

Abstract Image

Abstract Image

用于在鸡蛋生物反应器中生产重组蛋白的表达系统的评价。
鸡蛋由于其基因修饰而作为优秀的生物反应器而受到关注。然而,鸡蛋生物反应器的开发从生产体系的构建到产品的评价需要很长的时间。因此,在本研究中,我们建立了一个生产卵清蛋白(OVA)的鸡细胞系,并构建了一个快速评估生产系统的系统。首先,我们敲除鸡DF-1细胞中OVA基因座上游的EF1α启动子,以连续表达OVA。此外,我们分析并鉴定了OVA基因座上插入有用蛋白质基因的理想位置,以最大限度地提高重组蛋白质产量。我们发现敲除外显子1上游的EF1α启动子产生了OVA蛋白的最大产量。此外,我们发现将重组hFGF2 cDNA连接到OVA的5’侧时生产效率更高。因此,我们建立了表达OVA的细胞系和鸡蛋生物反应器中蛋白质的评估系统。我们的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。这篇文章受版权保护。保留所有权利。
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来源期刊
Biotechnology Journal
Biotechnology Journal Biochemistry, Genetics and Molecular Biology-Molecular Medicine
CiteScore
8.90
自引率
2.10%
发文量
123
审稿时长
1.5 months
期刊介绍: Biotechnology Journal (2019 Journal Citation Reports: 3.543) is fully comprehensive in its scope and publishes strictly peer-reviewed papers covering novel aspects and methods in all areas of biotechnology. Some issues are devoted to a special topic, providing the latest information on the most crucial areas of research and technological advances. In addition to these special issues, the journal welcomes unsolicited submissions for primary research articles, such as Research Articles, Rapid Communications and Biotech Methods. BTJ also welcomes proposals of Review Articles - please send in a brief outline of the article and the senior author''s CV to the editorial office. BTJ promotes a special emphasis on: Systems Biotechnology Synthetic Biology and Metabolic Engineering Nanobiotechnology and Biomaterials Tissue engineering, Regenerative Medicine and Stem cells Gene Editing, Gene therapy and Immunotherapy Omics technologies Industrial Biotechnology, Biopharmaceuticals and Biocatalysis Bioprocess engineering and Downstream processing Plant Biotechnology Biosafety, Biotech Ethics, Science Communication Methods and Advances.
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