Journal of Animal and Veterinary Advances
ISSN: Online 1993-601XISSN: Print 1680-5593
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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
Page: 251-256 | Received 21 Sep 2022, Published online: 21 Sep 2022
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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.
How to cite this article:
Jian Tu, Kezong Qi, Xiuheng Xue, Haixia Wang, Xueyan Wang, Kaisong Peng, Lingling Xie and Yanli Wu. Construction of Recombinant pPIC9K-AvBD2 and Expression of Chicken AvBD2
Mature Peptide in Pichia pastoris.
DOI: https://doi.org/10.36478/javaa.2014.251.256
URL: https://www.makhillpublications.co/view-article/1680-5593/javaa.2014.251.256