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Sensing mispaired thymines in DNA heteroduplexes using an electroactive osmium marker: towards electrochemical SNP probing
P. Kostečka, L. Havran, M. Bittová, H. Pivoňková, M. Fojta
Jazyk angličtina Země Německo
Typ dokumentu časopisecké články, práce podpořená grantem
NLK
ProQuest Central
od 2011-01-01 do Před 1 rokem
Medline Complete (EBSCOhost)
od 2003-01-01 do Před 1 rokem
Health & Medicine (ProQuest)
od 2011-01-01 do Před 1 rokem
- MeSH
- DNA sondy MeSH
- DNA chemie MeSH
- elektrochemie metody MeSH
- heteroduplexy nukleové kyseliny chemie MeSH
- jednonukleotidový polymorfismus MeSH
- osmium chemie MeSH
- sekvence nukleotidů MeSH
- thymin analýza MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A complex OsO(4), 2,2'-bipyridine (Os,bipy), has been used for electroactive labeling of biopolymers as well as for probing of nucleic acids and protein structure and interactions. In DNA, Os,bipy forms electrochemically active adducts with pyrimidine nucleobases, exhibiting highly selective modification of thymine residues in single-stranded DNA. Here, we show that modification of rare thymine residues (one thymine among several tens of unreactive purine bases) can easily be detected by means of a simple ex situ voltammetric analysis using carbon electrodes. Based on this remarkable sensitivity of detection, Os,bipy has been used as an electroactive probe for unpaired and/or mismatched thymine residues within DNA heteroduplexes. Site-specific chemical modification of the DNA with the Os,bipy has allowed a clear distinction between perfectly base-paired DNA homoduplexes and mismatched heteroduplexes, as well as discrimination among heteroduplexes containing one or two mispaired thymines, a single thymine insertion, or combination of a mispair and an insertion.
Faculty of Science Department of Chemistry Masaryk University
Institute of Biophysics Academy of Sciences of the Czech Republic
Citace poskytuje Crossref.org
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- $a A complex OsO(4), 2,2'-bipyridine (Os,bipy), has been used for electroactive labeling of biopolymers as well as for probing of nucleic acids and protein structure and interactions. In DNA, Os,bipy forms electrochemically active adducts with pyrimidine nucleobases, exhibiting highly selective modification of thymine residues in single-stranded DNA. Here, we show that modification of rare thymine residues (one thymine among several tens of unreactive purine bases) can easily be detected by means of a simple ex situ voltammetric analysis using carbon electrodes. Based on this remarkable sensitivity of detection, Os,bipy has been used as an electroactive probe for unpaired and/or mismatched thymine residues within DNA heteroduplexes. Site-specific chemical modification of the DNA with the Os,bipy has allowed a clear distinction between perfectly base-paired DNA homoduplexes and mismatched heteroduplexes, as well as discrimination among heteroduplexes containing one or two mispaired thymines, a single thymine insertion, or combination of a mispair and an insertion.
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