Abstract
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.
MeSH terms
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DNA / chemistry
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DNA Nucleotidylexotransferase / metabolism
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Deoxyuracil Nucleotides / chemical synthesis*
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Deoxyuracil Nucleotides / chemistry*
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Deoxyuracil Nucleotides / metabolism
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Deoxyuridine / analogs & derivatives
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Deoxyuridine / chemical synthesis*
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Energy Transfer
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Fluorescence
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Fluorescent Dyes / chemistry*
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Fluorescent Dyes / metabolism
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Humans
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Kinetics
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Molecular Structure
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Oligodeoxyribonucleotides / chemical synthesis*
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Oligodeoxyribonucleotides / chemistry
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Organometallic Compounds / chemical synthesis*
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Organometallic Compounds / chemistry*
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Organometallic Compounds / metabolism
Substances
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Deoxyuracil Nucleotides
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Fluorescent Dyes
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K-11-dUTP
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Oligodeoxyribonucleotides
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Organometallic Compounds
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deoxyuridine triphosphate
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europium(III) trisbipyridine cryptate
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DNA
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DNA Nucleotidylexotransferase
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Deoxyuridine