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. 2003 Aug;77(16):8621–8632. doi: 10.1128/JVI.77.16.8621-8632.2003

FIG. 1.

FIG. 1.

FIG. 1.

Incorporation of ddATP, CBV-TP, and d4T-TP by WT and mutant RTs. (A) Graphic representation of the cell-free system (PPT-57-PPT-19) used to monitor the incorporation of ddATP by WT and mutant RTs. The DNA template consists of 57 nucleotides designed to correspond to the PPT sequence of the HIV-1 genome. The primer was 5′ end labeled. Bold underlined letters indicate sites of chain termination. Dots define positions +1 and +4 in the reaction. (B) Dose-dependent incorporation of ddATP by WT and mutant RTs. Reactions were performed with increasing concentrations of ddATP (lanes 1 to 15). The concentration of dATP was kept at 1 μM, in order to allow incorporation of the analogue ddATP, which was employed at increasing concentrations (0, 0.1, 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 μM). The other dNTPs were used at 10 μM. A control reaction was performed without RT enzyme (lanes C). (C) Graphic representation of the cell-free system (PPT-57-PPT-18) used to monitor incorporation of CBV-TP. (D) Dose-dependent incorporation of CBV-TP by WT and mutant RTs. Reactions were performed as described for panel B, except that ddATP was replaced by CBV-TP (0, 0.1, 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 μM) and the concentration of dGTP was 1 μM.