NDLI: Zebrafish eggs used as bioreactors for the production of bioactive tilapia insulin-like growth factors

Content Provider	Springer Nature Link
Author	Hu, Shao Yang Liao, Chia Hsuan Lin, Yi Pei Li, Yen Hsing Gong, Hong Yi Lin, Gen Hwa Kawakami, Koichi Yang, Tzu Hsuan Wu, Jen Leih
Copyright Year	2010
Abstract	Multiple advantages-including the short generation time, large numbers of fertilized eggs, low cost of cultivation and easy maintenance favor the use of fish as bioreactors for the production of pharmaceutical proteins. In the present study, zebrafish eggs were used as bioreactors to produce mature tilapia insulin-like growth factors (IGFs) proteins using the oocyte-specific zona pellucida (zp3) promoter. The chimeric expression plasmids, pT2-ZP-tIGFs-IRES-hrGFP, in which hrGFP was used as reporter of tilapia IGFs expression, were designed to established Tg (ZP:tIGFs:hrGFP) transgenic lines for the expression of tilapia IGF-1 and IGF-2. Recombinant tilapia IGF-1 and IGF-2 were expressed as soluble forms in cytoplasm of fertilized eggs. The content level of tilapia IGF-1 and IGF-2 were 6.5 and 5.0% of the soluble protein, respectively. Using a simple Ni–NTA affinity chromatography purification process, 0.58 and 0.49 mg of purified tilapia IGF-1 and IGF-2 were obtained, respectively, from 650 fertilized eggs. The biological activity of the purified tilapia IGF-1 and IGF-2 was confirmed via a colorimetric bioassay to monitor the growth stimulation of zebrafish embryonic cells (ZF4), tilapia ovary cells (TO-2) and human osteosarcoma epithelial cells (U2OS). These results demonstrate that the use of zebrafish eggs as bioreactors is a promising approach for the production of biological recombinant proteins.
Starting Page	73
Ending Page	83
Page Count	11
File Format	PDF
ISSN	09628819
Journal	Transgenic Research
Volume Number	20
Issue Number	1
e-ISSN	15739368
Language	English
Publisher	Springer Netherlands
Publisher Date	2010-04-07
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Tilapia Insulin-like growth factors Bioreactor Fish eggs Zona pellucida Zebrafish Human Genetics Plant Sciences Animal Genetics and Genomics Plant Genetics & Genomics Molecular Medicine
Content Type	Text
Resource Type	Article
Subject	Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science

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