Figure 5.

(A) Gel autoradiogram illustrating representative RNase H inhibitory activity resulting from screening DONAS hairpin library members. A [³²P]-RNA:DNA hybrid was incubated with HIV-1 RT in the presence of varying amounts of hairpins (0–50 μM). (B) Inhibition assay of DNA synthesis catalyzed by (a) DNA-dependent DNA polymerase and (b) RNA-dependent DNA polymerase activities of HIV-1 RT. Unlabeled hairpin (80 μM) was pre-incubated with RT for 20 min at room temperature prior to initiating the polymerization reaction by adding the proper DNA or RNA template hybridized to a 5′-[³²P]-labeled primer in the presence of dNTPs (see Materials and Methods). The various lanes show full-length DNA products synthesized after 15 min in the absence (−) and presence (+) of unlabeled hairpins.