Abstract

THE CONSTRUCTION AND FUNCTIONAL STUDIES OF THE RECOMBINANT PROTEIN VEC-SOD CONTAINING MEMBRANE-PENETRATING PEPTIDE

Yi Wu, Zhuo-Ying Lin, Wen-Yu Yao, Jian-Zhou Chen and Ren-Wang Jiang*

ABSTRACT

Purpose: In order to enhance the cell membrane penetration ability of superoxide dismutase, SOD1 and the cell penetrating peptide VEC were fused in this study. Methods: The VEC-SOD1 expression plasmid was successfully constructed. The recombinant plasmid was transformed into Escherichia coli BL21(DE3). The induction agent IPTG was used to induce the expression of the fusion protein, and the purified protein was obtained by nickel column purification. After purification, the enzyme activity of the protein was detected by the WST-1 method, and the total antioxidant capacity in vitro was detected by the ABTS method and the cell membrane penetration ability was determined by Western Blot analysis. Results: The experimental results show that the VEC-SOD1 fusion protein has significantly increased transmembrane ability and strong antioxidant effect. Conclusion: The cell penetrating peptide VEC was successfully fused with SOD1, and VEC-SOD1 showed promising antioxidant activity and increased cell membrane penetration ability.

Keywords: VEC-SOD1; fusion protein; superoxide dismutase; cell penetrating peptide.