Evaluation of cell wall binding domain of Staphylococcus aureus autolysin as affinity reagent for bacteria and its application to bacterial detection.
J Biosci Bioeng
; 104(1): 55-61, 2007 Jul.
Article
en En
| MEDLINE
| ID: mdl-17697984
We evaluated the cell wall binding (CWB) domain of Staphylococcus aureus autolysin as an affinity reagent for bacteria. A fusion of CWB domain and green fluorescent protein (CWB-GFP) bound to S. aureus with a dissociation constant of 15 nM. CWB-GFP bound to a wide range of gram-positive bacteria, but not to most gram-negative bacteria. We suspected that the outer membrane of gram-negative bacteria inhibits the access of CWB-GFP to peptidoglycan layer. Indeed, CWB-GFP bound to gram-negative bacteria when they were treated with benzalkonium chloride. Because CWB-GFP bound to the bacterial peptidoglycan layer, it appeared to be an effective affinity reagent for bacteria and CWB fusion with reporter proteins could be applied to detect bacteria. We also constructed a fusion of CWB and luciferase, which can be used for the rapid detection of bacteria.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Staphylococcus aureus
/
Bacterias
/
Pared Celular
/
Técnicas Bacteriológicas
/
Proteínas Fluorescentes Verdes
/
N-Acetil Muramoil-L-Alanina Amidasa
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
J Biosci Bioeng
Asunto de la revista:
ENGENHARIA BIOMEDICA
/
MICROBIOLOGIA
Año:
2007
Tipo del documento:
Article
País de afiliación:
Japón
Pais de publicación:
Japón