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Biotechnology and applied biochemistry

The construction of a bifunctional fusion protein consisting of SEC2 and EGFP.


PMID 24397332

Abstract

The aim of this study was to construct a bifunctional fusion protein consisting of staphylococcal enterotoxin C2 (SEC2) and enhanced green fluorescent protein (EGFP). We inserted EGFP and SEC2 fragments into the pET-28a(+) vector to create the expression plasmid vector, pET-28a(+)-SEC2-EGFP, using a two-step method. After verification of the plasmid, successful isolation of the fusion protein, SEC2-EGFP, was achieved by Ni+-affinity chromatography. Fluorescence microscopy, methylthiazol tetrazolium, and flow cytometry assays demonstrated that the constructed fusion protein not only retained the fluorescence signal of EGFP but also exhibited SEC2 bioactivity. Therefore, SEC2-EGFP is a promising tool for the study of the detailed temporal and spatial distributions of SEC2 in cells. Future studies with this vector may help uncover novel therapeutic strategies to treat or manage SEC2-associated diseases and be a new clinical tool for exploiting SEC2 in immunotherapy.