FIG. 1.

Representative gel-shift analysis of the PF1 aptamer and various derivatives. Increasing amounts of HIV RT (as indicated above wells; note different amounts with different constructs) were mixed with 5′ ³²P end-labeled PF1 or other modified constructs (see Table 1). Samples were run on a native 6% polyacrylamide gel. Positions of shifted and unshifted material are indicated. For PF1-4G, G-quartets (G-quart, boxed lanes) were observed migrating above the unshifted DNA. A small portion of the material remained in the gel wells with some aptamers, but this occurred even when no RT was added, indicating it was not dependent on the added protein. In general, PF1 and the other derivatives tested did not form a single discrete band in gel-shift assays. A concentrated shifted band was observed just below the wells and a diffuse area of shifted material appeared underneath the band. This probably resulted from some complexes dissociating during the electrophoresis process.