Skip to main content
Figure 1 | AIDS Research and Therapy

Figure 1

From: HIV-1 reverse transcriptase mutations that confer decreased in vitro susceptibility to anti-RT DNA aptamer RT1t49 confer cross resistance to other anti-RT aptamers but not to standard RT inhibitors

Figure 1

RNase H cleavage of RNA.DNA hybrids by wild type (WT) and mutant RTs in the presence of a heparin challenge. A. Polymerase-dependent RNase H clevage. The substrate, as diagrammed at the top, consisted of a 142nt heteropolymeric RNA (thin line) annealed to a 30nt DNA primer (thick line). Arrows indicate the expected sites of cleavage. Reactions were performed in the absence of dNTPs and in the presence of a heparin trap. Control reactions were performed in which either no enzyme was added (C), or an RNAse H-defective mutant (E478Q) was added (RNase H-) (see Methods secion). Cleavage products were resolved on a denaturing 6% polyacrylamide gel. The sizes of the resultant radiolabeled products are represented to the left of the gel panels (including a minor product). B. RNA 5'-end-directed RNase H cleavage. The substrate was a 41nt heteropolymeric RNA annealed to a 47nt DNA template. Reaction conditions were otherwise identical to those in panel A, and are described under 'Materials and Methods' section. Cleavage products were resolved on a denaturing 12% polyacrylamide gel. The sizes of the resultant radiolabeled products are represented to the left of the gel panels.

Back to article page