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Base‐Modified DNA Labeled by [Ru(bpy) 3 ] 2+ and [Os(bpy) 3 ] 2+ Complexes: Construction by Polymerase Incorporation of Modified Nucleoside Triphosphates, Electrochemical and Luminescent Properties, and Applications
Author(s) -
Vrábel Milan,
Horáková Petra,
Pivoňková Hana,
Kalachova Lubica,
Černocká Hana,
Cahová Hana,
Pohl Radek,
Šebest Peter,
Havran Luděk,
Hocek Michal,
Fojta Miroslav
Publication year - 2009
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.200801538
Subject(s) - chemistry , dna , polymerase , nucleobase , redox , stereochemistry , nucleotide , primer (cosmetics) , ruthenium , nucleic acid , dna polymerase , nucleoside , linker , combinatorial chemistry , biochemistry , organic chemistry , computer science , gene , operating system , catalysis
Abstract Modified 2′‐deoxynucleoside triphosphates (dNTPs) bearing [Ru(bpy) 3 ] 2+ and [Os(bpy) 3 ] 2+ complexes attached via an acetylene linker to the 5‐position of pyrimidines (C and U) or to the 7‐position of 7‐deazapurines (7‐deaza‐A and 7‐deaza‐G) have been 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 with Ru 2+ or Os 2+ complexes. Square‐wave voltammetry could be efficiently used to detect these labeled nucleic acids by reversible oxidations of Ru 2+/3+ or Os 2+/3+ . The redox potentials of the Ru 2+ complexes (1.1–1.25 V) are very close to that of G oxidation (1.1 V), while the potentials of Os 2+ complexes (0.75 V) are sufficiently different to enable their independent detection. On the other hand, Ru 2+ ‐labeled DNA can be independently analyzed by luminescence. In combination with previously reported dNTPs bearing ferrocene, aminophenyl, and nitrophenyl tags, the Os‐labeled dATP has been successfully used for “multicolor” redox labeling of DNA and for DNA minisequencing.