Table of Contents Author Guidelines Submit a Manuscript
Journal of Biomedicine and Biotechnology
Volume 2005 (2005), Issue 4, Pages 374-384
Research article

Recombinant Expression of Pleurocidin cDNA Using the Pichia pastoris Expression System

1Department of Food Science, Center for Advanced Food Technology, and Institute of Coastal & Marine Sciences, Rutgers, The State University of New Jersey, New Brunswick 08901, NJ, USA
2Department of Oral Biology, New Jersey Dental School, University of Medicine & Dentistry of New Jersey (UMDNJ), 110 Bergen Street, Newark 07103-2495, NJ, USA

Received 27 September 2004; Revised 13 April 2005; Accepted 26 April 2005

Copyright © 2005 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


This research utilized the Pichia pastoris expression system for recombinant expression of cDNA of pleurocidin, a small (2.7 kd) antimicrobial peptide isolated from winter flounder (Pleuronectes americanus). The Pichia vector contains the alcohol oxidase gene promoter (AOX 1), which under the induction of methanol allows for expression of heterologous protein gene inserted downstream in the vector. Two strains of P pastoris were used as host cells, the wild type (P pastoris X-33a(mut+)) and the mutant (P pasatoris KM71Ha(muts)). Polymerase chain reaction (PCR) and DNA sequencing showed that pleurocidin cDNA was successfully integrated into the P pastoris genome. Reverse transcription (RT)-PCR showed that pleurocidin was transcribed by both Pichia host strains. Affinity chromatography, SDS-PAGE, and immunological techniques were used for purification and detection of recombinant peptide. Although there was strong evidence of transcription of pleurocidin cDNA, the Pichia system requires further optimization to obtain detectable levels of this small peptide.