RESUMO
5-Aminoallyl-2'-fluoro-dUTP, 5-aminoallyl-UTP, and N(6)-([6-aminohexyl]carbamoylmethyl)-ATP were systematically tested for their suitability for the systematic evolution of ligands by exponential enrichment (SELEX) process with the aim of introducing additional functionalities to RNA libraries. All three aminomodified nucleoside triphosphates proved to be compatible with the enzymatic steps required for SELEX and maintained strict Watson-Crick basepairing. Complementary RNA molecules modified with the two uridine analogues show a significantly increased melting temperature, whereas the introduction of N(6)-([6-aminohexyl]carbamoylmethyl)-ATP leads to a decreased T(m) and thus less stable basepairing. The chemical synthesis of 5-aminoallyl-2'-fluoro-dUTP is reported in detail.
Assuntos
Trifosfato de Adenosina/química , Análise de Sequência de RNA/métodos , Uridina Trifosfato/química , Trifosfato de Adenosina/genética , Animais , Sequência de Bases , Bovinos , Venenos de Crotalídeos , Crotalus , Biblioteca Gênica , RNA/análise , RNA/química , RNA/genética , Uridina Trifosfato/genéticaRESUMO
Building bridges: A feasible solid-phase synthesis of antiparallel n-, h-, and H-type DNA oligomers has been demonstrated. The oligomers possess a CH2 -bridged base-pair model that should be conformationally flexible, even after base pairing.