Figure 3.
Recombinant HIV-1 RT catalyzes reverse transcription and is augmented by recombinant RHA. The synthesis of (−)ssDNA was primed by a 5′-Cy3–labeled tRNALys3 annealed to the HIV-1 5′-UTR template, and reactions were collected and analyzed at the indicated time points. The RNA size marker is labeled with M at the top of the gels. A, visualization of the annealed RNA complexes on a native agarose gel. PBS segment and tRNALys3 (1 μm) were annealed by NC (30 μm) in the absence and presence of RHA (1 μm) at 37 °C. B, diagram of the 5′-Cy3–labeled tRNALys3 annealed to the HIV-1 5′-UTR RNA template. C, comparison of the tRNALys3 primer extension products, synthesized in the absence and presence of RHA (1 μm) and separated by 8% urea–PAGE. Representative gels are shown.