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Polymerase Synthesis and Restriction Enzyme Cleavage of DNA Containing 7‐Substituted 7‐Deazaguanine Nucleobases
Author(s) -
Mačková Michaela,
Boháčová Soňa,
Perlíková Pavla,
Poštová Slavětínská Lenka,
Hocek Michal
Publication year - 2015
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201500315
Subject(s) - nucleobase , restriction enzyme , dna polymerase , dna , chemistry , cleavage (geology) , enzyme , polymerase , dna clamp , biochemistry , stereochemistry , biology , polymerase chain reaction , gene , reverse transcriptase , paleontology , fracture (geology)
Abstract 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 (dG R TPs), 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 dG R TPs 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.