Zhen Zhang, Tao Ke, Yuling Zhou, Xiangdong Ma, Lixin Ma
College of Life Science, Hubei University, Wuhan 430062, China.
Sheng wu gong cheng xue bao = Chinese journal of biotechnology 2009 AugIn this study, we efficiently expressed the active antimicrobial peptide (CAD), which fused with the site-mutated coat protein (EDDIE) of the classical swine fever virus, in Escherichia coli. First, we obtained the e-cad fusion gene from the CAD gene and the EDDIE gene using overlapping PCR. Then to get the recombinant expression vector (pETED), the e-cad fusion gene was cloned into the pET30a vector by a site-directed homologous recombination technique. The EDDIE-CAD fusion protein expressed in E. coli as inclusion bodies, and its yield was more than 40% of total bacterial proteins. After renaturated in vitro and self-cleavage of the fusion protein, we obtained the antimicrobial peptide Cecropin AD. Antimicrobial experiments showed that the Cecropin AD efficiently inhibited the growth of G+ and G- bacteria, but it weakly inhibited the growth of Saccharomyces. This method provides an excellent way for high expression of antimicrobial peptides when fused with EDDIE.
Zhen Zhang, Tao Ke, Yuling Zhou, Xiangdong Ma, Lixin Ma. High expression of antimicrobial peptide Cecropin AD in Escherichia coli by fusion with EDDIE]. Sheng wu gong cheng xue bao = Chinese journal of biotechnology. 2009 Aug;25(8):1247-53
PMID: 19938464
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