Abstract
Human β-defensin-2 (hBD2) is A small cationic peptide with A broad range of antimicrobial activity. An E. coli cell-free system was employed to express the hBD2 fusion protein by using the hBD2 gene with 14 rare codons. The results showed that the expression level of trxA-hBD2 fusion protein was 0.35 mg/ml, which is the same as that obtained with A synthetic codon-optimized gene. By using another fusion partner (GFP), similar high-level expression was also achieved in this cell-free system. This meant that human beta-defensin-2 gene could be directly used to express hBD2 fusion protein efficiently in an E. coli cell-free system without the optimization of codons. The expression level of hBD2 fused with thioredoxin could be further improved up to 2.0 mg/ml by adopting A continuous exchange cell-free system. A simple one-stage affinity purification procedure was also developed to recover this fusion protein efficiently.
Keywords: Human beta-defensin-2, cell-free system, codon usage, fusion protein, antimicrobial peptide, protein purification
Protein & Peptide Letters
Title: High-Level Expression of Human β-Defensin-2 Gene with Rare Codons in E. coli Cell-Free System
Volume: 13 Issue: 2
Author(s): Haiqin Chen, Zhinan Xu, Naizheng Xu and Peilin Cen
Affiliation:
Keywords: Human beta-defensin-2, cell-free system, codon usage, fusion protein, antimicrobial peptide, protein purification
Abstract: Human β-defensin-2 (hBD2) is A small cationic peptide with A broad range of antimicrobial activity. An E. coli cell-free system was employed to express the hBD2 fusion protein by using the hBD2 gene with 14 rare codons. The results showed that the expression level of trxA-hBD2 fusion protein was 0.35 mg/ml, which is the same as that obtained with A synthetic codon-optimized gene. By using another fusion partner (GFP), similar high-level expression was also achieved in this cell-free system. This meant that human beta-defensin-2 gene could be directly used to express hBD2 fusion protein efficiently in an E. coli cell-free system without the optimization of codons. The expression level of hBD2 fused with thioredoxin could be further improved up to 2.0 mg/ml by adopting A continuous exchange cell-free system. A simple one-stage affinity purification procedure was also developed to recover this fusion protein efficiently.
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Cite this article as:
Chen Haiqin, Xu Zhinan, Xu Naizheng and Cen Peilin, High-Level Expression of Human β-Defensin-2 Gene with Rare Codons in E. coli Cell-Free System, Protein & Peptide Letters 2006; 13 (2) . https://dx.doi.org/10.2174/092986606775101724
DOI https://dx.doi.org/10.2174/092986606775101724 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
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