High-level expression of human beta-defensin-2 gene with rare codons in E. coli cell-free system |
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Authors: | Chen Haiqin Xu Zhinan Cen Peilin |
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Affiliation: | Institute of Bioengineering, Department of Chemical Engineering and Bioengineering, Zhejiang University, Hangzhou 310027, The People's Republic of China. |
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Abstract: | Human beta-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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