[High expression of antimicrobial peptide Cecropin AD in Escherichia coli by fusion with EDDIE].
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.
Zhang
Zhen
Z
College of Life Science, Hubei University, Wuhan 430062, China.
Ke
Tao
T
Zhou
Yuling
Y
Ma
Xiangdong
X
Ma
Lixin
L
D000890:Anti-Infective Agents / Q000378:metabolism; D036022:Capsid Proteins / Q000235:genetics* / Q000378:metabolism; D054807:Cecropins / Q000096:biosynthesis* / Q000235:genetics; D006692:Classical Swine Fever Virus / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D009154:Mutation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology
D000890:Anti-Infective Agents; D036022:Capsid Proteins; D054807:Cecropins; D011993:Recombinant Fusion Proteins