Efficient production of soluble human beta-defensin-3-4 fusion proteins in Escherichia coli cell-free system

被引:10
作者
Chen, Haiqin
Fan, Limei
Xu, Zhinan [1 ]
Yin, Xiufei
Cen, Peilin
机构
[1] Zhejiang Univ, Dept Chem & Biochem Engn, Hangzhou 310027, Peoples R China
[2] Zhejiang Univ City Coll, Hangzhou 310015, Peoples R China
[3] Zhejiang Yangshengtang Nat Med Inst, Hangzhou 310007, Peoples R China
关键词
human beta-defensin-3; human beta-defensin-4; antimicrobial peptide; cell-free system; soluble expression; GFP;
D O I
10.1016/j.procbio.2006.10.002
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Human beta-defensin-3 and 4 (HBD-3-4) are two low molecular weight cationic peptides with three conserved cysteine disulfide bonds, and exhibit a broad range of antimicrobial activity and do not acquire any microbial resistance. In order to produce these uneasily detectable, degradable and toxic polypeptide efficiently, an alternative approach based on the Escherichia coli cell-free biosynthesis system was proposed. The polypeptide of interest is synthesized as a fusion protein linked to trxA or green fluorescent protein (GFP) through a cleavable spacer. With batch mode operation, significant amount of hBD3-4 fused with trxA or GFP can be expressed in this cell-free system, and the product is soluble and stable. Furthermore, the GFP moiety provides directly visuable and quantitative monitoring of the polypeptide synthesis. This work will be helpful to rapid and visuable expression of other similar defensins using in vitro cell-free system. (c) 2006 Published by Elsevier Ltd.
引用
收藏
页码:423 / 428
页数:6
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