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Journal of Animal and Veterinary Advances
Year: 2014  |  Volume: 13  |  Issue: 4  |  Page No.: 251 - 256

Construction of Recombinant pPIC9K-AvBD2 and Expression of Chicken AvBD2 Mature Peptide in Pichia pastoris

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.

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