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Cleavage of Functionalized DNA Containing 5-Modified Pyrimidines by Type II Restriction Endonucleases
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SYSNO ASEP 0360993 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Cleavage of Functionalized DNA Containing 5-Modified Pyrimidines by Type II Restriction Endonucleases Author(s) Macíčková-Cahová, Hana (UOCHB-X) RID, ORCID
Pohl, Radek (UOCHB-X) RID, ORCID
Hocek, Michal (UOCHB-X) RID, ORCIDNumber of authors 3 Source Title Chembiochem. - : Wiley - ISSN 1439-4227
Roč. 12, č. 3 (2011), s. 431-438Number of pages 8 s. Language eng - English Country DE - Germany Keywords base-modified DNA ; DNA cleavage ; DNA polymerases ; nucleosides ; restriction endonucleases Subject RIV CC - Organic Chemistry R&D Projects LC512 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) GA203/09/0317 GA ČR - Czech Science Foundation (CSF) CEZ AV0Z40550506 - UOCHB-X (2005-2011) UT WOS 000287718700016 DOI 10.1002/cbic.201000644 Annotation A series of six pyrimidine modified dNTPs: 5-ethynyl- 5-phenyl- and 5-(3-nitrophenyl)-deoxycitidine and -deoxyuridine triphosphates were prepared and incorporated by primer extension using Vent (exo-) polymerase to specific DNA sequences within or next to the recognition sequences of selected restriction endonucleases. Then the cleavage of these pyrimidine-modified DNA sequences by 13 restriction enzymes was studied. While the presence of any modified C within the target sequence completely prevented any restriction cleavage, most enzymes tolerated the presence of 5-ethynylU and two of them even the presence of 5-phenyl- and 5-(3-nitrophenyl)U. Modifications outside the recognition sequence were tolerated apart from phenyl derivatives with PvuII enzyme. 5-EthynylC was used for protection of the recognition sequence from cleavage in presence of the second unmodified copy of the same sequence that was cleaved. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Viktorie Chládková, Tel.: 232 002 434 Year of Publishing 2012
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