Expression of fluorescent tagged recombinant erythroferrone protein

Min Min Than; Jetsada Ruangsuriya; Somdet Srichairatanakool; Chairat Uthaipibull; Min Min Than

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Expression of fluorescent tagged recombinant erythroferrone protein

Author: Min Min THAN ¹ ; Jetsada RUANGSURIYA ¹ ; Somdet SRICHAIRATANAKOOL ¹ ; Chairat UTHAIPIBULL ² ; Min Min THAN ³
Author Information

1. Department of Biochemistry, Faculty of Medicine, Chiang Mai University
2. Protein-Ligand Engineering and Molecular Biology Laboratory, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency
3. Department of Biochemistry, University of Medicine
Publication Type:Journal Article
Keywords: Erythroferrone; HEK293T cell; Hepcidin; Iron; Recombinant protein
From:Asian Pacific Journal of Tropical Biomedicine 2018;8(7):360-364
CountryChina
Language:Chinese
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.