Heterologous production of porcine antimicrobial peptide PR-39 in Escherichia coli using SUMO and intein fusion systems

In this study, we presented two different expression systems for the production of PR-39 in E. coli; one in fusion with Intein-Chitin binding domain (CBD) and another in fusion with SUMO accompanied by polyhistidine affinity tag. Both were cloned in the NdeI-XhoI sites of pET-17b and transformed to E. coli BL21 (DE3) pLysS. Recombinant bacteria were cultured and induced with 0.4 mM IPTG at 30 °C. Expression and purification of target proteins were confirmed by Tricine- SDS-PAGE and dot blot analysis. Recovery of 250 μg PR-39/L from SUMO fusion system and 280 μg PR-39/L from the intein fusion system was achieved.
Source: Protein Expression and Purification - Category: Biochemistry Source Type: research