Europium cryptate labeled deoxyuridine-triphosphate analog: synthesis and enzymatic incorporation.
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.
Alpha-Bazin
B
B
CIS biointernational/DIVT/Research and New technologies, Bagnols/Cèze, France.
Bazin
H
H
Guillemer
S
S
Sauvaigo
S
S
Mathis
G
G
D004247:DNA / Q000737:chemistry; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism; D003856:Deoxyuracil Nucleotides / Q000138:chemical synthesis* / Q000737:chemistry* / Q000378:metabolism; D003857:Deoxyuridine / Q000031:analogs & derivatives / Q000138:chemical synthesis*; D004735:Energy Transfer; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D015394:Molecular Structure; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D009942:Organometallic Compounds / Q000138:chemical synthesis* / Q000737:chemistry* / Q000378:metabolism
D003856:Deoxyuracil Nucleotides; D005456:Fluorescent Dyes; C420282:K-11-dUTP; D009838:Oligodeoxyribonucleotides; D009942:Organometallic Compounds; C027078:deoxyuridine triphosphate; C070734:europium(III) trisbipyridine cryptate; D004247:DNA; D004253:DNA Nucleotidylexotransferase; D003857:Deoxyuridine