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

07:00 EST 11th January 2020 | BioPortfolio

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

About half a century after antibiotics discovery, multi-antibiotic-resistant bacteria posed a new challenge to medicine. Attempts to discover new antibiotics have drawn the attention to Antimicrobial Peptides (AMPs). The rapid growth, besides its known genetic and manipulation systems, makes E.coli the preferred host system for production of recombinant proteins on an industrial scale. To produce AMPs in E.coli, the application of fusion-tags with the aim of stability, solubility, and prevention of antimicrobial activity is one of the best practices in this regard. 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.

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This article was published in the following journal.

Name: Protein expression and purification
ISSN: 1096-0279
Pages: 105568

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