Cloning and expression of visfatin and screening of oligopeptides binding with visfatin

Int J Clin Exp Med. 2014 Dec 15;7(12):4828-34. eCollection 2014.

Abstract

Purpose: In the present study, we constructed an efficient expression system for visfatin and identified the peptide sequences which binding to visfatin.

Methods and results: The fusion protein was identified by SDS-PAGE and Western blot. The yield of visfatin was 300 mg/L culture medium with optimal conditions. The recombinant visfatin binds with insulin receptor with a dose-dependent effect. All of the 24 sequence identified by ELISA were able to bind to visfatin specifically. Among the 24 DNA sequences, there were 8 clones of AAKTPTE, 4 clones of ATTVPAS and 4 clones of MSLQQEH.

Conclusion: The sequence of AA(X)TPT(X) was the most frequently existed sequence in all of sequences analyzed, which suggests that AA(X)TPT(X) is likely to be the essential motif in peptides which visfatin could bind with.

Keywords: Visfatin; adipokine; fusion expression; gene engineering; phage display.