Non-radioactive labeling and detection of nucleic acids. IV. Synthesis and properties of digoxigenin-modified 2'-deoxyuridine-5'-triphosphates and a photoactivatable analog of digoxigenin (photodigoxigenin).
The chemical syntheses of novel digoxigenin-derivatized compounds are described which are modified substrates for enzymatically or photochemically non-radioactive digoxigenin labeling of nucleic acids. Various activated digoxigenin-haptens are coupled to 5-aminoallyl-substituted 2'-deoxyuridine-5'-triphosphate. This results in digoxigenin-modified nucleoside triphosphates of variable spacer lengths (Dig-[4]-dUTP/Dig-[11]-dUTP/Dig-[16]-dUTP) which can be used as substrates for enzymatic labeling of DNA with digoxigenin-haptens by Klenow enzyme-catalysed random-primed synthesis. In addition the synthesis of N-[4-azidobenzoyl]-N'-[(3-O-digoxigeninyl)methylcarbonyl)]-1 ,8-diamino- 3,6-dioxaoctane (photodigoxigenin), a photoactivatable analog of digoxigenin, is described which can be applied for photolabeling of DNA and RNA with digoxigenin-haptens leaving the nucleic acid molecules intact.
Mühlegger
K
K
Boehringer Mannheim GmbH, Biochemisches Forschungszentrum Penzberg, Tutzing.
Huber
E
E
von der Eltz
H
H
Rüger
R
R
Kessler
C
C
D004247:DNA / Q000032:analysis / Q000378:metabolism; D003856:Deoxyuracil Nucleotides / Q000138:chemical synthesis; D004076:Digoxigenin / Q000031:analogs & derivatives / Q000378:metabolism; D009696:Nucleic Acids / Q000032:analysis* / Q000378:metabolism; D009711:Nucleotides / Q000138:chemical synthesis; D014544:Uridine Triphosphate / Q000138:chemical synthesis
D003856:Deoxyuracil Nucleotides; D009696:Nucleic Acids; D009711:Nucleotides; D004247:DNA; D004076:Digoxigenin; D014544:Uridine Triphosphate