High expression of antimicrobial peptide Cecropin AD in Escherichia coli by fusion with EDDIE.
- Author:
Zhen ZHANG
1
;
Tao KE
;
Yuling ZHOU
;
Xiangdong MA
;
Lixin MA
Author Information
1. College of Life Science, Hubei University, Wuhan 430062, China.
- Publication Type:Journal Article
- MeSH:
Anti-Infective Agents;
metabolism;
Capsid Proteins;
genetics;
metabolism;
Cecropins;
biosynthesis;
genetics;
Classical swine fever virus;
genetics;
metabolism;
Escherichia coli;
genetics;
metabolism;
Genetic Vectors;
genetics;
Mutation;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
pharmacology
- From:
Chinese Journal of Biotechnology
2009;25(8):1247-1253
- CountryChina
- Language:Chinese
-
Abstract:
In 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.
