RESUMO
C8-Arylamine-dG and C8-arylamine-dA adducts have been prepared using palladium cross-coupling chemistry. These adducts were subsequently converted into the corresponding 5'-O-DMTr-C8-arylamine-3'-O-phosphoramidites and then used for the automated synthesis of different site-specifically modified oligonucleotides. These "damaged" oligonucleotides have been characterized by ESI-MS, UV thermal stability assays, and circular dichroism, and they have been used in EcoRI assays as well as in primer extension studies using various DNA polymerases.
Assuntos
Adutos de DNA/síntese química , Dano ao DNA , DNA Polimerase Dirigida por DNA/efeitos dos fármacos , Oligodesoxirribonucleotídeos/síntese química , Purinas/química , Desoxirribonuclease EcoRI , Técnicas de Amplificação de Ácido Nucleico , Oligodesoxirribonucleotídeos/química , Compostos Organofosforados , Paládio , Análise EspectralRESUMO
The synthesis of C8- and N(6)-adducts of 2'- deoxyadenosine with carcinogenic aromatic amines is described. Furthermore, the adducts were converted into their corresponding phosphoramidites. These were applied to solid phase DNA synthesis to obtain the site-specifically modified oligonucleotides that were characterized and investigated regarding melting temperature, circular dichroism spectra and in some cases by enzymatic degradation assays.
