Expression of fluorescent tagged recombinant erythroferrone protein
Asian Pacific Journal of Tropical Biomedicine
;
(12): 360-364, 2018.
Article
in Chinese
| WPRIM
| ID: wpr-700138
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 DH5α 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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Asian Pacific Journal of Tropical Biomedicine
Year:
2018
Type:
Article
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