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. 2000 Apr 4;97(8):3832–3837. doi: 10.1073/pnas.070471597

Figure 4.

Figure 4

DNA repair synthesis in response to a cyPu lesion. (a) The plasmid M13mp18 cyclo-dA is shown diagrammatically. Eight BstNI restriction sites and the three AlwI restriction sites are indicated. One AlwI site (underlined) overlaps the region containing the cyPu residue. Unique PvuI, HindIII, and XhoI restriction enzyme sites are also indicated. (b) Autoradiograph after denaturing 14% PAGE, demonstrating DNA repair synthesis in the region containing the cyPu lesion. Plasmid DNA was incubated with HeLa cell extracts and radioactively labeled deoxynucleoside triphosphates (see Materials and Methods) and subsequently digested with BstNI before electrophoresis. Lanes 1–4, Pt-GTG substrate as positive control; lanes 5–8, control DNA without any lesion; lanes 9–12, 5′R-cyclo-dA substrate; lanes 13–16, 5′S-cyclo-dA substrate. M, size markers as in Fig. 2.