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Base-modified DNA labeled by Ru2+(bpy)3 and Os2+(bpy)3 complexes. Construction by polymerase incorporation of modified nucleoside triphosphates, electrochemical and luminescent properties and applications
Authors: Vrábel Milan | Horáková Petra | Pivoňková Hana | Kalachova Lubica | Černocká Hana | Cahová Hana | Pohl Radek | Šebest Peter | Havran Luděk | Hocek Michal | Fojta Miroslav
Year: 2009
Type of publication: článek v odborném periodiku
Name of source: Chemistry - A European Journal
Publisher name: Wiley-VCH
Place: Weinheim
Page from-to: 1144-1154
Titles:
Language Name Abstract Keywords
cze Byly připraveny modifikované 2'-deoxynukleosid trifosfáty (dNTPs) nesoucí Ru2+(bpy)3 and Os2+(bpy)3 complexy připojené acetylenovým linkrem k pozici 5 pyrimidinů (C a U) nebo k pozici 7 7-deazapurinů (7-deaza-A a 7-deaza-G). Pomocí DNA polymeráz byly získány DNA řetězce značené ve specifických pozicích komplexy Ru2+ nebo Os2+. Elektrochemickou detekcí (SWV) takto značených nukleových kyselin byly získány specifické signály odpovídající těmto komplexům. Komplex Ru2+ byl detekován také pomocí luminiscence. V kombinaci s dříve publikovanými dNTP nesoucími ferocen, amino- a nitro-skupinu bylo dATP značené komplexem Os2+ úspěšně využito při vícebarevném značení DNA a pro minisekvencování DNA.
eng Base-modified DNA labeled by Ru2+(bpy)3 and Os2+(bpy)3 complexes. Construction by polymerase incorporation of modified nucleoside triphosphates, electrochemical and luminescent properties and applications Modified 2'-deoxynucleoside triphosphates (dNTPs) bearing Ru2+(bpy)3 and Os2+(bpy)3 complexes attached via acetylene linker to position 5 of pyrimidines (C and U) or to position 7 of 7-deazapurines (7-deaza-A and 7-deaza-G) were prepared in one step by aqueous cross-couplings of halogenated dNTPs with the corresponding terminal acetylenes. Polymerase incorporation by primer extension using Vent (exo-) or Pwo polymerases gave DNA labeled in specific positions by Ru2+ or Os2+ complexes. The redox potentials of the Ru2+ complexes are very close to G oxidation, while the potentials of Os2+ complexes differ significantly to enable independent DNA labelling. On the other hand, Ru2+ labeled DNA can be independently detected by luminescence. In combination with previously reported dNTPs bearing ferrocene, amino- and nitrophenyl tags, the Os-labeled dATP was successfully used for "multicolor" redox labelling of DNA and for DNA minisequencing. modified nucleoside triphosphates;DNA labelling;DNA sequencing;square-wave voltammetry

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