Sensing mispaired thymines in DNA heteroduplexes using an electroactive osmium marker: towards electrochemical SNP probing

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Publikace nespadá pod Fakultu sportovních studií, ale pod Přírodovědeckou fakultu. Oficiální stránka publikace je na webu muni.cz.
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KOSTEČKA Pavel HAVRAN Luděk BITTOVÁ Miroslava PIVOŇKOVÁ Hana FOJTA Miroslav

Rok publikování 2011
Druh Článek v odborném periodiku
Časopis / Zdroj Analytical and Bioanalytical Chemistry
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
www http://www.springerlink.com/content/074k47jthp76x862/
Obor Analytická chemie, separace
Klíčová slova Single-base mismatch; Nucleobase insertion; Osmium tetroxide; Carbon electrodes; Voltammetry; Chemical probe; Electroactive label
Popis A complex OsO4, 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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