Journal: Nucleic Acids Research
Article Title: Divergent evolution of opposite base specificity and single-stranded DNA activity in animal and plant AP endonucleases
doi: 10.1093/nar/gkae1297
Figure Lengend Snippet: Orphan base preference in plant and bacterial AP endonucleases. ( A ) Opposite base preference in a single sequence context. APE1 (0.02 nM), ARP (40 nM), ExoIII (0.06 nM) or EndoIV (0.3 nM) were incubated with DNA substrates (20 nM) containing a native AP site opposite G (orange), A (green), T (yellow) or C (blue) in the sequence context 5′-A X G-3′ ( X = AP site). ( B ) Opposite base preference in different sequence contexts. DNA substrates (20 nM) containing a native AP site opposite G (orange) or C (blue) in four different sequence contexts were incubated with APE1 (0.01 nM), ARP (40 nM), ExoIII (0.06 nM) or EndoIV (0.9 nM). The reaction products were stabilized with NaBH 4 , separated by denaturing PAGE and detected by fluorescence scanning for quantification. Data are the mean and standard error from two independent experiments. Asterisks indicate significant differences in a two-tailed Student’s t -test (* P < 0.05; ** P < 0.01; *** P < 0.001).
Article Snippet: Human APE1 complementary DNA (cDNA) was synthetized by codon optimization for expression in bacteria (GenScript).
Techniques: Sequencing, Incubation, Fluorescence, Two Tailed Test