Authors : Jian Tu, Kezong Qi, Xiuheng Xue, Haixia Wang, Xueyan Wang, Kaisong Peng, Lingling Xie and Yanli Wu
Abstract: Avian β-Defensin-2 (AvBD2) is an antimicrobial peptide that plays a significant role in the innate immunity of chickens. In this research, the expression vector pPIC9K-AvBD2 was established and expressed in Pichia pastoris. AvBD2 cDNA was cloned from total RNA of chicken lung by Transcriptase-Polymerase Chain Reaction (RT-PCR). The AvBD2 mature peptide gene was amplified from and cloned into a pPIC9K plasmid. Then the recombinant expression vector pPIC9K-AvBD2 was transformed into P. pastoris GS115 to express the fusion protein. The result showed that a recombinant secretion expression vector pPIC9K-AvBD2 was successfully constructed and a fusion protein about 3.9 kDa chicken AvBD2 was expressed. Densitometric scanning analysis demonstrated that the expressed protein exaccounted for about 63.7% of the total secreted bacterial protein. The expression products had antimicrobial activity to Escherichia coli and Staphylococcus aureus in the Assay of Agar Diffusion Method.
Jian Tu, Kezong Qi, Xiuheng Xue, Haixia Wang, Xueyan Wang, Kaisong Peng, Lingling Xie and Yanli Wu, 2014. Construction of Recombinant pPIC9K-AvBD2 and Expression of Chicken AvBD2 Mature Peptide in Pichia pastoris. Journal of Animal and Veterinary Advances, 13: 251-256.