Cleavage of functionalized DNA containing 5-modified pyrimidines by type II restriction endonucleases.
Chembiochem
; 12(3): 431-8, 2011 Feb 11.
Article
em En
| MEDLINE
| ID: mdl-21290545
ABSTRACT
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 with Vent (exo-)polymerase to specific DNA sequences within or next to the recognition sequences of selected restriction endonucleases. The cleavage of these pyrimidine-modified DNA sequences by 13 restriction enzymes was then studied. Whereas 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 except in the case of phenyl derivatives with the PvuII enzyme. 5-EthynylC was used for protection of the recognition sequence from cleavage in the presence of the second unmodified copy of the same sequence that was cleaved.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
DNA
/
Desoxirribonucleases de Sítio Específico do Tipo II
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article