Expression of EGFP-Arg7 Fusion Protein and Its Transduction Activity into HeLa Cells

Objective To construct an expression vector of pET-28a-EGFP-Arg7 and to investigate its transduction activity into cells.Methods EGFP-Arg7 gene was amplified by PCR and cloned into prokaryotic expression vector pET-28a.The constructed recombinant plasmid pET-28a-EGFP-Arg7 was transformed to E.coli BL21(DE3) for the expression under the induction of IPTG.The expressed protein was purified by Ni2+-NTA affinity chromatography and its transduction activity into the HeLa cells was detected under the fluorescence microscope.Results Both endonuclease restriction analysis and sequence analysis proved that the recombinant plasmid pET-28a-EGFP-Arg7 was constructed correctly.The recombinant fusion protein was expressed in a soluble form,with a purity of more than 90 % after the purification.It showed strong green fluorescence into the HeLa cells under the fluorescence microscope.Conclusion EGFP-Arg7 fusion protein has a good transduction activity and can take the linked protein through the HeLa cells.