RESUMO
The synthesis of an europium tris-bipyridine cryptate labeled 2'-deoxyuridine-5 '-triphosphate analog (K-11-dUTP) is described. This labeled triphosphate was incorporated into DNA through enzymatic reactions with terminal transferase and DNA polymerases. The enzymatic reactions were monitored by TRACE (Time Resolved Amplification of Cryptate Emission), a homogeneous method using Fluorescence Resonance Energy Transfer (FRET) from an europium cryptate as donor to a modified allophycocyanine as acceptor.
Assuntos
Nucleotídeos de Desoxiuracil/química , Nucleotídeos de Desoxiuracil/síntese química , Desoxiuridina/síntese química , Corantes Fluorescentes/química , Oligodesoxirribonucleotídeos/síntese química , Compostos Organometálicos/química , Compostos Organometálicos/síntese química , DNA/química , DNA Nucleotidilexotransferase/metabolismo , Nucleotídeos de Desoxiuracil/metabolismo , Desoxiuridina/análogos & derivados , Transferência de Energia , Fluorescência , Corantes Fluorescentes/metabolismo , Humanos , Cinética , Estrutura Molecular , Oligodesoxirribonucleotídeos/química , Compostos Organometálicos/metabolismoRESUMO
A noncompetitive N-methyl-D-aspartate (NMDA) antagonist, MK-801 (0.5 to 2.0 mM), inhibits rabies virus infection in rat primary cortical neurons, whereas the competitive NMDA antagonist AP5 has no effect. The results suggest that MK-801-mediated inhibition of rabies virus replication, although selective, is not operating through the high-affinity binding site mechanism.
