Heterologous expression and purification of dermaseptin S4 fusion in Escherichia coli and recovery of biological activity.
Prep Biochem Biotechnol
; 44(6): 598-607, 2014.
Article
em En
| MEDLINE
| ID: mdl-24499364
ABSTRACT
Heterologous expression of dermaseptin S4 (DS4), which has cytolytic activity in vitro against a broad spectrum of pathogenic microorganisms, was examined in Escherichia coli. The plasmid pGEX-4T-1, encoding DS4 fused with glutathione S-transferase (GST), was constructed and cloned into the E. coli strain BL21 (DE3). The fusion protein was overexpressed in this strain after induction with isopropyl-beta-D-thiogalactopyranoside (IPTG) and purified to homogeneity using GST affinity chromatography. To recover biologically active DS4, the purified fusion protein was cleaved using thrombin protease; the liberated DS4 was shown to be bactericidally active against an indicator strain. Since it is less expensive to obtain such a peptide biologically, in this study, we report for the first time a method to express purify DS4 in E. coli using a GST fusion system.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteínas Recombinantes de Fusão
/
Expressão Gênica
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Peptídeos Catiônicos Antimicrobianos
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Proteínas de Anfíbios
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Escherichia coli
Limite:
Animals
Idioma:
En
Revista:
Prep Biochem Biotechnol
Assunto da revista:
BIOQUIMICA
/
BIOTECNOLOGIA
Ano de publicação:
2014
Tipo de documento:
Article
País de afiliação:
China