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by
Jian Tu |
Total Records (
2 ) for
Jian Tu |
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Kezong Qi
,
Liu Miao
,
Kaisong Peng
,
Jian Tu
and
Xiuheng Xue
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Chicken Beta Defensin-8 (AvBD8) is an antibacterial peptide
that plays significant roles in innate immunity. The aim of this study was to
investigate the expression of Chicken avian AvBD8 mature peptide gene in Lactococcus
lactis (L. lactis) system. In this study, the AvBD8 gene was
cloned and sequenced from chicken liver by Reverse Transcriptase-Polymerase
Chain Reaction (RT-PCR). Then, the AvBD8 mature peptide gene was amplified and
cloned into the expression vector pNZ8048. The result showed that the recombinant
vector pNZ8048-AvBD8 that verified by colony PCR, double digestion and DNA sequencing
was transformed into the competent cell L. lactis NZ9000 by electroporation.
The expression of the recombinant protein AvBD8 was confirmed by tricine SDS-PAGE
analysis. The present findings suggest that L. lactis could act as a
host for the production of avian beta defensins for further study. |
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Jian Tu
,
Kezong Qi
,
Xiuheng Xue
,
Haixia Wang
,
Xueyan Wang
,
Kaisong Peng
,
Lingling Xie
and
Yanli Wu
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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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