Fig. 5.
RSC is dispensable for SSA recombination. (A) Diagram showing SSA in tNS1379. The 205-bp ura3 direct repeats, the HO recognition site, the locations of BglII sites (arrows), and the probe (black bar with asterisk) used to detect SSA products in the Southern blot assay are shown. (B) Percent survival was determined by dividing the number of colonies on a YEP-galactose plate with that in YEPD. Each experimental point represents the average of three independent experiments ± the standard deviation. (C) SSA products in rsc mutants were detected by Southern blot hybridization. Genomic DNA digested with BglII was separated by agarose gel electrophoresis and subjected to Southern blot hybridization with 32P-labeled probes. One anneals to the 3′ region of the ura3 sequences (as indicated in panel A), and the other anneals to the RAD1 gene at chromosome XVI. U, signal represents uncut fragment; C, signal resulting from the HO cleavage; R, signal from recombination; H, signal from the RAD1 locus used as a control. (D) Percent SSA repair was determined by normalizing the amount of SSA products with that of the control RAD1 signal and plotted as a function of time of HO expression. Results obtained with the wild-type RSC strain (WT; tNS1379) and the rsc2Δ, rsc7Δ, and rad59Δ mutants are shown. Each point represents the average of three independent experiments ± the standard deviation.