RESUMEN
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.
Asunto(s)
Nucleótidos de Desoxiuracil/química , Nucleótidos de Desoxiuracil/síntesis química , Desoxiuridina/síntesis química , Colorantes Fluorescentes/química , Oligodesoxirribonucleótidos/síntesis química , Compuestos Organometálicos/química , Compuestos Organometálicos/síntesis química , ADN/química , ADN Nucleotidilexotransferasa/metabolismo , Nucleótidos de Desoxiuracil/metabolismo , Desoxiuridina/análogos & derivados , Transferencia de Energía , Fluorescencia , Colorantes Fluorescentes/metabolismo , Humanos , Cinética , Estructura Molecular , Oligodesoxirribonucleótidos/química , Compuestos Organometálicos/metabolismoRESUMEN
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.