Extended Data Fig. 5: The strand-displacement activity analysis of the Pol1A and Pol1B.
From: PEN1 catalyses RNA primer removal during plastid DNA replication in maize
a, Schematic of the linear gapped substrates used in strand-displacement assays, with the resulting products depicted in each corresponding diagram. The magenta lines indicate the ribonucleotides. The green star represents the /6FAMdT/ modification. To prevent end-binding by the Pol1A and Pol1B, all the unlabeled ends were blocked by the streptavidin. The black balls represent the streptavidin modification. b, Coupled primer extension and ligation mediated by Pol1A, Pol1B, and LIG1. The displaced primer is 5 nt longer than the template strand, so the ligated product (65 nt) can be distinguished from the displaced product (60 nt), which is the same size as the template strand. The black balls represent the streptavidin modification. M, molecular Marker (values indicate the number of nucleotides). The green stars represent the /6FAMdT/ modification. Reactions were terminated at 15 min and analyzed on a high-resolution denaturing polyacrylamide gel. Three replicates, consistent results.
