Informace o publikaci

Protected 5-(hydroxymethyl) uracil nucleotides bearing visible-light photocleavable groups as building blocks for polymerase synthesis of photocaged DNA

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BOHACOVA Sona LUDVÍKOVÁ Lucie SLAVETINSKA POSTOVA Lenka VANIKOVA Zuzana KLÁN Petr HOCEK Michal

Rok publikování 2018
Druh Článek v odborném periodiku
Časopis / Zdroj ORGANIC & BIOMOLECULAR CHEMISTRY
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
www http://pubs.rsc.org/en/Content/ArticleLanding/2018/OB/C8OB00160J#!divAbstract
Doi http://dx.doi.org/10.1039/c8ob00160j
Klíčová slova STEM-CELL DNA; RESTRICTION ENDONUCLEASES; MAMMALIAN-CELLS; RNA-POLYMERASE; CHEMICAL-MODIFICATIONS; PHOTOCHEMICAL CONTROL; GENE-EXPRESSION; NUCLEIC-ACIDS; MODIFIED BASE; TRANSCRIPTION
Popis Nucleosides, nucleotides and 2'-deoxyribonucleoside triphosphates (dNTPs) containing 5-(hydroxy-methyl) uracil protected with photocleavable groups (2-nitrobenzyl-, 6-nitropiperonyl or 9-anthrylmethyl) were prepared and tested as building blocks for the polymerase synthesis of photocaged oligonucleotides and DNA. Photodeprotection (photorelease) reactions were studied in detail on model nucleoside mono-phosphates and their photoreaction quantum yields were determined. Photocaged dNTPs were then tested and used as substrates for DNA polymerases in primer extension or PCR. DNA probes containing photocaged or free 5-hydroxymethylU in the recognition sequence of restriction endonucleases were prepared and used for the study of photorelease of caged DNA by UV or visible light at different wavelengths. The nitropiperonyl-protected nucleotide was found to be a superior building block because the corresponding dNTP is a good substrate for DNA polymerases, and the protecting group is efficiently cleavable by irradiation by UV or visible light (up to 425 nm).
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