TY - JOUR T1 - Construction of Recombinant pPIC9K-AvBD2 and Expression of Chicken AvBD2 Mature Peptide in Pichia pastoris AU - Qi, Kezong AU - Tu, Jian AU - Xue, Xiuheng AU - Wang, Haixia AU - Wang, Xueyan AU - Peng, Kaisong AU - Xie, Lingling AU - Wu, Yanli JO - Journal of Animal and Veterinary Advances VL - 13 IS - 4 SP - 251 EP - 256 PY - 2014 DA - 2001/08/19 SN - 1680-5593 DO - javaa.2014.251.256 UR - https://makhillpublications.co/view-article.php?doi=javaa.2014.251.256 KW - AvBD2 mature peptide KW -P. pastoris KW -cloning KW -expression KW -antibacterial activity AB - 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. ER -