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. 2000 Aug 15;97(17):9537–9542. doi: 10.1073/pnas.97.17.9537

Figure 2.

Figure 2

DSBs at ura3∷URA3-arg4 in normal and hybrid diploids. Southern blots contain DNA from diploids with a hemizygous ura3∷URA3-arg4 insert and homozygous for the rad50-K81I mutation, containing either two S. cerevisiae chromosomes V (homo) or an S. cerevisiae and an S. carlsbergensis chromosome V (homoeo). DNA samples from premeiotic cells (mit, lanes 1, 4, 7, and 10) and cells 5 and 6 h after induction of meiosis (mei, 5 h, lanes 2, 5, 8, and 11; mei, 6 h, lanes 3, 6, 9, and 12) were restriction enzyme-digested, displayed on 0.5% agarose gels, transferred to membranes, and hybridized with radioactive probe (36). (a) Restriction enzyme, SmaI; probe, pBR322 HindIII–BamHI fragment. (b) Restriction enzyme, EcoRI; probe, pBR322 PstI–EcoRI fragment. Horizontal arrows, meiosis-induced DSBs at ARG4 (DSB ARG4) and in pBR322 sequences on either side of ARG4 sequences (DSB left and DSB right). Numbers in parentheses are ratios of DSB frequencies for each site (6-h samples, DSB-homoeo/DSB-homo; n.d., not determined). In the insert diagrams, thick lines, pBR322; hatched boxes, URA3; open boxes, ARG4; thin lines, Ty sequences; thick gray lines, probe. A Ty element inserted in URA3 provides one of the EcoRI sites in B (T.-C. Wu and M.L., unpublished data).