Expressing of porcine β-defensin-2 mature peptide in the yeast

Abstract:

Abstract: [Objective] Porcine β-defensin-2 mature peptide gene fragments were correctly integrated into genome chromosome of the yeast KM71, which was stable Pichia pastoris strains expressing porcine β-defensin-2 mature peptide. Porcine β-defensin-2 mature peptide was expressed successfully. [Method] Considering the biased codon usage of Pichia pastoris, we designed three primers and used PCR technology to get the gene fragment of pBD-2 mature peptide. We obtained the recombinant vectors of pPIC9K-pBD-2 and pPIC9k-GST-pBD-2. The recombinant plasmids linearized were transformed into yeast KM71 by electroporation. By changing the parameters of expression environment, screening yeast clones induced by methanol expressed the Porcine β-defensin-2 mature peptide finally. [Result] GST-Porcine β-defensin-2 and Porcine β-defensin-2 gene fragments were successfully integrated into genome chromosome of the yeast KM71, We finally constructed the recombinant yeast; Protein GST-Porcine β-defensin-2 and Porcine β-defensin-2 were expressed successfully. The peptide of Porcine β-defensin-2 can restrain the Salmonella choleraesuis (C500-). [Conclusion] We got the recombinant yeast which can express porcine β-defensin-2 mature peptide. This study is an exploration to express Porcine β-defensin-2 in eukaryotic cells, It established foundation for the subsequent study of large-scale expression of Porcine β-defensin-2 mature peptide.