Incorporation of 4-thiothymidine into DNA by the Klenow fragment and HIV-1 reverse transcriptase.
The 5'-triphosphate of 4-thiothymidine (4S-TTP) is an excellent substrate for the Klenow fragment of Escherichia coli DNA polymerase 1 and HIV-1 reverse transcriptase with values of k(cat)/Km within a factor of approximately 3 of those for TTP. A large UV change (deltaepsilon= -9770 M(-1)cm(-1) at 340 nm) associated with incorporation of 4S-TMP into nucleic acid duplexes makes possible a rapid, continuous spectrophotometric assay of the reaction progress.
Rao
T V
TV
National Institutes of Health, Bethesda, MD 20892, USA.
Haber
M T
MT
Sayer
J M
JM
Jerina
D M
DM
D020029:Base Pairing / Q000187:drug effects; D004247:DNA / Q000096:biosynthesis* / Q000737:chemistry; D004256:DNA Polymerase I / Q000378:metabolism*; D017931:DNA Primers; D004926:Escherichia coli / Q000201:enzymology; D054303:HIV Reverse Transcriptase / Q000378:metabolism*; D007700:Kinetics; D013056:Spectrophotometry, Ultraviolet; D013698:Templates, Genetic; D013936:Thymidine / Q000031:analogs & derivatives* / Q000737:chemistry
C414668:4-thiothymidine; D017931:DNA Primers; D004247:DNA; D054303:HIV Reverse Transcriptase; D004256:DNA Polymerase I; D013936:Thymidine