[Derivatives of ddUTP, modified at the 5-position of uridine, as substrate terminators of reverse transcriptase. Hydrolysis of oligonucleotides, terminated by these analogs, by phosphodiesterase I].
Synthesis of 2',3'-dideoxyuridine 5'-triphosphate analogues with fluorescent and biotin residues at C5 of uracil base was carried out. The substrate properties of these analogues were studied with AMV, M-MLV, and HIV reverse transcriptase. All 5-derivatives studied were shown to be incorporated into the 3'-terminus of oligonucleotide. The stability of oligodeoxyribonucleotides terminated with ddUTP analogues modified at the 5-position of the pyrimidine residue to the exonuclease action of phosphodiesterase I and Klenow enzyme was more than 1000 times higher than that of nonterminated oligonucleotides.
D'iachenko
L B
LB
Chenchik
A A
AA
Lukin
M A
MA
Aleksandrova
L A
LA
Kraevskiĭ
A A
AA
Bibilashvili
R Sh
RSh
D009189:Avian Myeloblastosis Virus / Q000201:enzymology; D001483:Base Sequence; D004256:DNA Polymerase I / Q000378:metabolism; D054306:Dideoxynucleotides; D006678:HIV / Q000201:enzymology; D006868:Hydrolysis; D008324:Mammary Tumor Virus, Mouse / Q000201:enzymology; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D013379:Substrate Specificity; D013728:Terminator Regions, Genetic; D014500:Uracil Nucleotides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D014529:Uridine / Q000737:chemistry*
D054306:Dideoxynucleotides; D009838:Oligodeoxyribonucleotides; D014500:Uracil Nucleotides; C063034:2',3'-dideoxyuridine-5'-triphosphate; D012194:RNA-Directed DNA Polymerase; D004256:DNA Polymerase I; D010727:Phosphoric Diester Hydrolases; D043264:Phosphodiesterase I; D014529:Uridine