Microarray of DNA–protein complexes on poly-3-hydroxybutyrate surface for pathogen detection
Copyright policy )Microarray of DNA–protein complexes on poly-3-hydroxybutyrate surface for pathogen detection
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.
- University of Queensland Australia
- University of Queensland Australia
- Yonsei University Health System Korea (Republic of)
- Yonsei University Medical Library Korea (Republic of)
- Korea Advanced Institute of Science and Technology Korea (Republic of)
Acinetobacter baumannii, 570, Hydroxybutyrates/chemistry*, Surface Properties, Polyesters, Recombinant Fusion Proteins, Protein microarray, Hydroxybutyrates, Pathogen detection, Polymerase Chain Reaction, Sensitivity and Specificity, Pseudomonas aeruginosa/genetics, Substrate Specificity, Immobilized/metabolism, Klebsiella pneumoniae/genetics, Oligonucleotide Array Sequence Analysis/instrumentation, Acinetobacter baumannii/genetics, Escherichia coli, Polyesters/chemistry*, Carboxylic Ester Hydrolases/metabolism, Escherichia coli/isolation & purification, DNA chip, Oligonucleotide Array Sequence Analysis, Pseudomonas aeruginosa/isolation & purification*, Recombinant Fusion Proteins/genetics, Substrate-binding domain, Binding Sites, 500, Recombinant Fusion Proteins/chemistry, Enzymes, Immobilized, Enzymes, DNA-Binding Proteins, Escherichia coli/genetics, Klebsiella pneumoniae, DNA Probes/chemistry, Acinetobacter baumannii/isolation & purification*, Oligonucleotide Array Sequence Analysis/methods*, Pseudomonas aeruginosa, PHB depolymerase, Poly-3-hydroxybutyrate, Klebsiella pneumoniae/isolation & purification*, DNA Probes, Carboxylic Ester Hydrolases, DNA-Binding Proteins/genetics, DNA-Binding Proteins/chemistry
Acinetobacter baumannii, 570, Hydroxybutyrates/chemistry*, Surface Properties, Polyesters, Recombinant Fusion Proteins, Protein microarray, Hydroxybutyrates, Pathogen detection, Polymerase Chain Reaction, Sensitivity and Specificity, Pseudomonas aeruginosa/genetics, Substrate Specificity, Immobilized/metabolism, Klebsiella pneumoniae/genetics, Oligonucleotide Array Sequence Analysis/instrumentation, Acinetobacter baumannii/genetics, Escherichia coli, Polyesters/chemistry*, Carboxylic Ester Hydrolases/metabolism, Escherichia coli/isolation & purification, DNA chip, Oligonucleotide Array Sequence Analysis, Pseudomonas aeruginosa/isolation & purification*, Recombinant Fusion Proteins/genetics, Substrate-binding domain, Binding Sites, 500, Recombinant Fusion Proteins/chemistry, Enzymes, Immobilized, Enzymes, DNA-Binding Proteins, Escherichia coli/genetics, Klebsiella pneumoniae, DNA Probes/chemistry, Acinetobacter baumannii/isolation & purification*, Oligonucleotide Array Sequence Analysis/methods*, Pseudomonas aeruginosa, PHB depolymerase, Poly-3-hydroxybutyrate, Klebsiella pneumoniae/isolation & purification*, DNA Probes, Carboxylic Ester Hydrolases, DNA-Binding Proteins/genetics, DNA-Binding Proteins/chemistry
selected citations These citations are derived from selected sources.This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).12 popularity This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.Average influence This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).Average impulse This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.Average
Citations provided by BIP!Popularity provided by BIP!