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

Autor: Kajihara R; Laboratory of Immunobiology, Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima, Hiroshima, Japan., Ezaki R; Laboratory of Immunobiology, Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima, Hiroshima, Japan., Watanabe T; Laboratory of Immunobiology, Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima, Hiroshima, Japan., Ichikawa K; 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., Matsuzaki M; Laboratory of Immunobiology, Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima, Hiroshima, Japan., Horiuchi H; 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.
Jazyk: angličtina
Zdroj: Biotechnology journal [Biotechnol J] 2024 Jan; Vol. 19 (1), pp. e2300316. Date of Electronic Publication: 2023 Oct 27.
DOI: 10.1002/biot.202300316
Abstrakt: 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.
(© 2023 Wiley-VCH GmbH.)
Databáze: MEDLINE
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