Polymerase Synthesis and Restriction Enzyme Cleavage of DNA Containing 7-Substituted 7-Deazaguanine Nucleobases

0451292 - ÚOCHB 2016 RIV DE eng J - Journal Article
Mačková, Michaela - Boháčová, Soňa - Perlíková, Pavla - Poštová Slavětínská, Lenka - Hocek, Michal
Polymerase Synthesis and Restriction Enzyme Cleavage of DNA Containing 7-Substituted 7-Deazaguanine Nucleobases.
Chembiochem. Roč. 16, č. 15 (2015), s. 2225-2236. ISSN 1439-4227. E-ISSN 1439-7633
R&D Projects: GA ČR GA14-04289S
Institutional support: RVO:61388963
Keywords : DNA * nucleotides * polymerases * pyrrolopyrimidines
Subject RIV: CC - Organic Chemistry
Impact factor: 2.850, year: 2015

Previous studies of polymerase synthesis of base-modified DNAs and their cleavage by restriction enzymes have mostly related only to 5-substituted pyrimidine and 7-substituted 7-deazaadenine nucleotides. Here we report the synthesis of a series of 7-substituted 7-deazaguanine 2'-deoxyribonucleoside 5'-O-triphosphates (dGRTPs), their use as substrates for polymerase synthesis of modified DNA and the influence of the modification on their cleavage by type II restriction endonucleases (REs). The dGRTPs were generally good substrates for polymerases but the PCR products could not be visualised on agarose gels by intercalator staining, due to fluorescence quenching. The presence of 7-substituted 7-deazaguanine residues in recognition sequences of REs in most cases completely blocked the cleavage.
Permanent Link: http://hdl.handle.net/11104/0252511