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| Fusion Expression and Purification of Human Beta Defensin-3 in E.coli |
| ZHU Jun-ping1, LI Gai-rui1, DU Qi-ke3, GUO Zhong-min2, LU Jia-hai1 |
1. School of Public Health, Sun Yat-sen University, Key Laboratory of Tropical Disease Control(Sun Yat-sen University), Ministry of Education, Guangdong Provincial Research Center for Severe Infectious Disease Prevention and Control Technology, Guangzhou 510080, China;
2. Experiment Animal Center, Sun Yat-sen University, Guangzhou 510080, China;
3. Good Earth Group, Animal Peptide Bio-Engineering Center of Shandong Province, Caoxian 274400, China |
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Abstract Antimicrobial peptides (AMPs) which are produced by most living organisms have shown robust activity against a wide variety of pathogens, including drug-resistant bacteria. They are important components of the innate and adaptive system. Human beta-defensin-3 is an endogenous, cysteine-rich antimicrobial peptide that resistants to high salt and contributes more powerful to host defense against multi-resistant bacteria compared with other beta-defensins. There is a great need for a less expensive and increasing efficiency peptide-production platform.This research aims at expressing human beta-defensin-3 and a novel sequence human beta-defensin-3i based on E. coli codon preference in E.coli BLR(DE3) via pET-EI plasmid vector. HβD-3i were 12 times higher yields than HβD-3. All the peptides, expressed as fusion protein, were isolated from the protein debris by the method called Inverse Transition Cycling(ITC).Fully reduced peptides that were purified exhibited expected antimicrobial activity that towarded to Staphylococus aureus was stronger than Salmonella.It is an attractive way to express human beta-defensin-3 via pET-EI prokaryotic plasmid vector. This approach described here is a low-cost, convenient and potential way for large-scale human beta-defensin-3 production.
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Received: 09 May 2013
Published: 25 November 2013
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