Fig 5.

Short annealed segment between target DNAs accelerates strand exchange. Strand exchange assays were performed with 1 μM Trx fusion of the N-terminal portion of ZfL2-1 ORF1p (Trx-N-ORF1p) at 37°C. (A to E and I to L) Schematic diagrams of the strand exchange assays. Oligonucleotides used in the assays are indicated by blue and red lines. Their sequences are shown in Table 2. One strand of each initial duplex was labeled by ³²P at its 5′ end (asterisks). (F and M) The initial rates of the strand exchange reactions (A to E and I to L, respectively). Five independent experiments of each strand exchange assay were performed, and the averages and standard deviations are shown. Statistical analysis was performed using Student's t test; ***, P < 0.001. (G and H) Strand exchange reactions between the duplex (co33-[³²P]o29) and increasing excess amounts of the single-stranded DNA (o29 in panel G or o33 in panel H) were induced by Trx-N-ORF1p, and their initial rates were calculated. Five independent experiments were performed for each concentration, and the averages and standard deviations are shown.