Construction Of Recombinant Plasmid PPIC9K-Hepcidin And Secret Expression And Characterization Of Human Hepcidin Growth Factor In Pichia Pastoris

Hepcidin is a kind of antimicrobial peptide with abundant cysteine. As indicated by researchers, it probably is the key substance which could control the iron absorbability of small intestine as well as accommodate the iron homeostasis in vivo.In this paper, amplifying Hepcidin from synthetical pMD18-T-Hepcidin by PCR, the gene fragment was inserted into the Pichia pastoris expression vector pPIC9K containing AOX1 promotor and a secreting signal peptides. The recombinant expression plasmid pPIC9K-Hepcidin was constructed and transformed into DH5a. After confirmed by DNA sequence analysis, this recombinant plasmid DNA was transformed into KM71. The multiple copy insert transformants were screened, fermented in flasks and induced by 0.5% methanol. The expression level of the protein of interest was detected by Tricine-SDS-PAGE, as well as the antimicrobial activity was verified by the way of the experiment of antimicrobial circles.