[Reverse transcriptase of the human immunodeficiency virus: isolation and substrate specificity].
Human immunodeficiency virus (HIV-I) reverse transcriptase was expressed in E. coli and purified to homogeneity (E. coli strain RRI (pRC-RT, pRK 248cIts)). We have investigated the substrate properties toward to DNA synthesis, catalyzed by this enzyme, of some nucleoside-5'-triphosphate analogues, previously studied in the same reactions, catalyzed by AMV and M-MLV reverse transcriptases. We have investigated substrate properties of new analogues of 2',3'-dideoxy-2',3'-didehydro- and 2',3'-dideoxytubercidin-5'-triphosphates. We have compared the relative efficiency of incorporation of different analogues tested in the DNA chain. It has been shown that expressed and purified HIV reverse transcriptase had the same specificity to analogues of 2'-deoxyribonucleoside-5'-triphosphates as was described for reverse transcriptases and natural HIV reverse transcriptase as well. These properties allow to apply the expressed HIV reverse transcriptase in different model systems.
Rozovskaia
T A
TA
Belogurov
A A
AA
Lukin
M A
MA
Chernov
D N
DN
Kukhanova
M K
MK
Bibilashvili
R Sh
RSh
D001483:Base Sequence; D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D004277:DNA, Single-Stranded; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D054303:HIV Reverse Transcriptase; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009711:Nucleotides / Q000737:chemistry / Q000378:metabolism*; D010957:Plasmids; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D013379:Substrate Specificity
D004277:DNA, Single-Stranded; D009711:Nucleotides; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase