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Microarray of DNA-protein complexes on poly-3-hydroxybutyrate surface for pathogen detection.

Authors
 Tae Jung Park  ;  Seung Min Yoo  ;  Ki Chang Keum  ;  Sang Yup Lee 
Citation
 ANALYTICAL AND BIOANALYTICAL CHEMISTRY, Vol.393(6-7) : 1639-1647, 2009 
Journal Title
 ANALYTICAL AND BIOANALYTICAL CHEMISTRY 
ISSN
 1618-2642 
Issue Date
2009
MeSH
Acinetobacter baumannii/genetics ; Acinetobacter baumannii/isolation & purification* ; Binding Sites ; Carboxylic Ester Hydrolases/metabolism ; DNA Probes/chemistry ; DNA-Binding Proteins/chemistry ; DNA-Binding Proteins/genetics ; Enzymes, Immobilized/metabolism ; Escherichia coli/genetics ; Escherichia coli/isolation & purification ; Hydroxybutyrates/chemistry* ; Klebsiella pneumoniae/genetics ; Klebsiella pneumoniae/isolation & purification* ; Oligonucleotide Array Sequence Analysis/instrumentation ; Oligonucleotide Array Sequence Analysis/methods* ; Polyesters/chemistry* ; Polymerase Chain Reaction ; Pseudomonas aeruginosa/genetics ; Pseudomonas aeruginosa/isolation & purification* ; Recombinant Fusion Proteins/chemistry ; Recombinant Fusion Proteins/genetics ; Sensitivity and Specificity ; Substrate Specificity ; Surface Properties
Keywords
Protein microarray ; DNA chip ; Poly-3-hydroxybutyrate ; Pathogen detection ; PHB depolymerase ; Substrate-binding domain
Abstract
A novel strategy was developed for the specific immobilization of DNA probes on poly-3-hydroxybutyrate (PHB) surface by using the substrate-binding domain (SBD) of PHB depolymerase as an active binding motif. To demonstrate whether this method can be used for the detection of clinical pathogens, the pathogen-specific biotin-labeled DNA probes were immobilized via core streptavidin (cSA) fused to the SBD. The pathogen-specific 15-mer oligonucleotide probes were designed for four model pathogens, while the target DNAs were prepared by PCR using universal primers. The complex of pathogen-specific probes and cSA-SBD fusion protein was immobilized on the PHB-coated slide by microspotting. This DNA-protein complex microarray was able to successfully diagnose Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Furthermore, the specific pathogens could be diagnosed in the presence of other microorganisms. Thus, the DNA-protein complex microarray platform technology employing PHB and the SBD reported here can be widely used for the detection of DNA-DNA and DNA-biomolecule interactions without synthetic or chemical modification of biomolecules or solid surface.
Full Text
http://link.springer.com/article/10.1007%2Fs00216-008-2574-y
DOI
10.1007/s00216-008-2574-y
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Radiation Oncology (방사선종양학교실) > 1. Journal Papers
Yonsei Authors
Keum, Ki Chang(금기창) ORCID logo https://orcid.org/0000-0003-4123-7998
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/105318
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