標題: Construction and expression of rabbit neutrophil peptide-1 gene in Escherichia coli
作者: Sun, Yu Ling
Kuan, Tang Ching
Lin, Yi Juain
Chou, Yu Chi
Lin, Chih Sheng
生物科技學系
Department of Biological Science and Technology
關鍵字: Antimicrobial activity;Fusion protein;Rabbit neutrophil peptide-1
公開日期: 1-六月-2010
摘要: Rabbit neutrophil peptide-1 (NP-1) is a prototypic rabbit alpha-defensin with a broad antimicrobial spectrum. The coding sequence of NP-1 was amplified and cloned into pET-31b(+) to construct an expression vector, pET31-NP1, which was transformed into E. coli BLR(DE3) pLysS for expressing the fusion NP-1 protein (fNP-1). The fNP-1 is downstream of a ketosteroid isomerase (KSI) and upstream of a (His)(6)-Tag, as KSI-NP1-His(6). The optimal condition, cultivation in enriched LB medium and induction with 0.5 mM IPTG for 6 h, was determined for fNP-1 production. The fNP-1 was purified by Ni-NTA resin and cleaved by cyanogen bromide to release matured NP-1 peptide. The matured NP-1 peptide showed significant antimicrobial activities against clinical bacteria, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus. The application of this expression approach represents a potential method to produce NP-1 by fusion protein expressed in E. coli without cytotoxicity.
URI: http://dx.doi.org/10.1007/s13213-010-0046-z
http://hdl.handle.net/11536/5287
ISSN: 1590-4261
DOI: 10.1007/s13213-010-0046-z
期刊: ANNALS OF MICROBIOLOGY
Volume: 60
Issue: 2
起始頁: 329
結束頁: 334
顯示於類別:期刊論文


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