Cloning, expression, and purification of insecticidal protein Pr596 from locust pathogen Serratia marcescens HR-3.
Curr Microbiol
; 55(3): 228-33, 2007 Sep.
Article
en En
| MEDLINE
| ID: mdl-17657528
A novel insecticidal protein (Pr596) produced by Serratia marcescens HR-3 was found be a metalloprotease and responsible for insecticidal activity toward locusts. Two pairs of primers were designed to amplify Pr596, a putative open reading frame (ORF) by similarity search and the N-terminal amino-acid sequence of insecticidal protein. The results revealed that the ORF consisted of 1464 nucleotides encoding a protein of 487 amino-acid residues. Pr596 was cloned into expression vector pET32a(+) and was expressed in Escherichia coli BL21 (DE3)/pLysS strain with isopropyl-beta-D-thiogalactopyranoside induction. The Pr596 was found to be highly expressed as inclusion bodies by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Pr596 inclusion bodies were isolated and subjected to Ni-NTA His Bind Resins (Pharmacia, Germany). Pr596 purified and refolded was revealed by SDS-PAGE and had proteolytic activity and insecticidal activity. Results suggested that there is a potential to develop this protein to be used as an alternative locus control agent.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Serratia marcescens
/
Proteínas Bacterianas
/
Saltamontes
Límite:
Animals
Idioma:
En
Revista:
Curr Microbiol
Año:
2007
Tipo del documento:
Article
Pais de publicación:
Estados Unidos