High-Level Expression of Human β-Defensin-2 Gene with Rare Codons in E. coli Cell-Free System

Authors: Chen, Haiqin; Xu, Zhinan; Xu, Naizheng; Cen, Peilin

Source: Protein and Peptide Letters, Volume 13, Number 2, February 2006 , pp. 155-161(7)

Publisher: Bentham Science Publishers

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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

Document Type: Research article

DOI: http://dx.doi.org/10.2174/092986606775101724

Affiliations: 1: Institute of Bioengineering, Department of Chemical Engineering and Bioengineering, Zhejiang University, Hangzhou 310027, The People's Republic of China.

Publication date: 2006-02-01

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Protein & Peptide Letters publishes short papers in all important aspects of protein and peptide research, including structural studies, recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, drug design etc. Manuscripts must have a significant element of novelty, timeliness and urgency that merit rapid publication. Reports of crystallisation, and preliminary structure determinations of biologically important proteins are acceptable. Purely theoretical papers are also acceptable provided they provide new insight into the principles of protein/peptide structure and function.

High-Level Expression of Human β-Defensin-2 Gene with Rare Codons in E. coli Cell-Free System

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