Fig. 5.

Effects of templates bearing HPMPC and HPMPA on DNA synthesis catalyzed by HIV-1 RT. Templates containing dCMP or HPMPC (top panel) or containing dAMP or HPMPA (bottom panel) were prepared and annealed to the indicated ³²P-labeled primers. These primers terminate one nucleotide prior to (P10 and P17), at (P13 and P18), or one nucleotide after (P14 and P19) the site of drug incorporation (marked with an “X”). The primer-template pairs were incubated with 200 μM dNTPs and 50 nM HIV-1 RT for 5 min at 37°C. The reaction products were recovered with magnetic beads and analyzed by phosphorimager analysis after separation on a 10% polyacrylamide gel. Lanes marked “Primer” show the position of each primer on the gel, and dideoxy sequencing reaction size markers are shown in lanes 10 to 13 of each gel.