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. Author manuscript; available in PMC: 2016 Mar 17.
Published in final edited form as: Nature. 2015 Aug 31;525(7569):345–350. doi: 10.1038/nature14887

Figure 4. Leading strands abut lagging strands of the opposing replisome during termination.

Figure 4

A. Cartoon illustrating the leading and lagging strands released by Nb.BtsI and Nb.BbvCI nicking enzymes. Primers JDO111 (purple arrow) and JDO110 (pink arrow) generated the sequencing ladders in (B) and (C), respectively.

B. LacR block-IPTG release was performed on p[lacOx12]. Termination intermediates were digested with Nb.BbvCI to liberate leftward leading strands, which were separated alongside a sequencing ladder on a denaturing polyacrylamide gel and visualized by autoradiography. Prominent leading strand products are highlighted (green symbols), and their sizes, measured relative to the Nb.BtsI site, are indicated.

C. Same samples as in (B) were digested with Nb.BtsI to liberate rightward lagging strands. The size of prominent lagging strand products (orange symbols), measured relative to the Nb.BtsI site, are indicated.

D. Schematic of the mapped leading (B) and lagging (C) strands.