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Expression of fluorescent tagged recombinant erythroferrone protein
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Metadata
Document Title
Expression of fluorescent tagged recombinant erythroferrone protein
Author
Than M.M., Ruangsuriya J., Uthaipibull C., Srichairatanakool S.
Name from Authors Collection
Affiliations
Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai, 50200, Thailand; Protein-Ligand Engineering and Molecular Biology Laboratory, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Thailand Science Park, Pathum Thani, Thailand; Department of Biochemistry, University of Medicine, Mandalay, Myanmar
Type
Article
Source Title
Asian Pacific Journal of Tropical Biomedicine
ISSN
22211691
Year
2018
Volume
8
Issue
7
Page
360-364
Open Access
Gold
Publisher
Wolters Kluwer Medknow Publications
DOI
10.4103/2221-1691.237079
Abstract
Objective: To produce fluorescent tagged recombinant erythroferrone protein (ERFE-eGFP) for laboratory investigations. Methods: Erythroferrone (ERFE) gene was fused to green fluorescent protein (eGFP) gene and cloned in a pSecTag2Hygro plasmid. The constructed plasmid was amplified in Escherichia coli DH5 α and the eGFP-fused ERFE (ERFE-eGFP) protein was expressed in human embryonic kidney (HEK293T) cell line. Results: The plasmid constructed from colony C6 contained ERFE-eGFP with the correct restriction sizes of 4.2 kb and expressed secretory ERFE-eGFP fusion protein (approximately size of 75 kDa) in HEK293T cell line. Conclusions: ERFE-eGFP recombinant protein is successfully expressed as a secretory functional protein and could be sensitively detected using fluorometry. This fusion protein might benefit future applications for localization of cellular ERFE receptors and competitive immunoassay of ERFE concentration. © 2018 by the Asian Pacific Journal of Tropical Biomedicine.
Keyword
erythroferrone | HEK293T cell | Hepcidin | Iron | Recombinant protein
License
CC BY-NC-SA
Rights
Wolters Kluwer Medknow Publications
Publication Source
Scopus