Kezong  Qi, Jian  Tu, Xiuheng  Xue, Haixia  Wang, Xueyan  Wang, Kaisong  Peng, Lingling  Xie, Yanli  Wu, 
    Construction of Recombinant pPIC9K-AvBD2 and Expression of Chicken AvBD2 
  Mature Peptide in <I>Pichia pastoris</I>,
    Journal of Animal and Veterinary Advances,
    Volume 13,Issue 4,
    2014,
    Pages 251-256,
    ISSN 1680-5593,
    javaa.2014.251.256,
    (https://makhillpublications.co/view-article.php?doi=javaa.2014.251.256)
    Abstract: Avian &#946;-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 <I>Pichia pastoris</I>. 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 <I>P. pastoris</I> 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 <I>Escherichia coli</I> and <I>Staphylococcus aureus</I> in the Assay of Agar Diffusion Method.
    Keywords: AvBD2 mature peptide;P. pastoris;cloning;expression;antibacterial activity