The Vecotor Construction, Expression And Preparation Of Hepcidin In Escherichia Coli

Hepcidin is a low-molecular-weight, highly disulfide bonded peptide relevant to small intestine iron absorption and body iron homeostasis. Like other cysteine-rich antimicrobial peptides, hepcidin also exhibits obvious antibacterial and antifungal activity. In this paper we express 25 aa human hepcidin as fusion proteins in E. coli and develop a preparation route of recombinant hepcidin.According to codon preference in E. coli and amino acid sequence of hepcidin, the gene coding for hepcidin was synthesized which was combined a encoding sequence of enterokinase cleavage site at its 5' end. Firstly we constructed pGEX-hpc. Although about 50% of GST-hepcidin fusion protein expressed from pGEX-hpc was soluble after induction with 0.1 mM IPTG at 25 ℃, it could not be absorbed by Glutathione -Sepharose. On the basis of pGEX-hpc the plasmid pET-hpc was constructed. The proportion of hepcidin in His-hepcidin was about 26.6%, and the fusion protein contained an N-terminal 6>