High expression of antimicrobial peptide Cecropin AD in Escherichia coli by fusion with EDDIE / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 1247-1253, 2009.
Article
in Chinese
| WPRIM
| ID: wpr-296931
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Recombinant Fusion Proteins
/
Capsid Proteins
/
Escherichia coli
/
Cecropins
/
Genetic Vectors
/
Genetics
/
Classical Swine Fever Virus
/
Metabolism
/
Anti-Infective Agents
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2009
Type:
Article
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