CHEF and Southern hybridization analysis of translocants unbalanced for Rdh54 and Rad54 amount.

<p>A) A scheme showing the bridge between chromosome VIII and XV and the new hybrid chromosome (Translocant = T). The chromosomal positions of the probes and of the translocation breakpoints (bold and underlined) are reported. B) CHEF (left) and Southern hybridization (right), with a probe against kanamycin, of the translocants in <i>RDH54</i> and <i>RAD54</i> deletants. Lanes from left to right: 1, San1ΔMSH2::Kan (it indicates chromosome XV); in <i>rdh54Δ/rdh54Δ</i> background: 2, cl4; 3, cl7; 4, cl16; 5, cl23n; 6, cl23*; 7, cl30; 8, cl47; in CRAD54 background: 9, cl1; 10, cl4; in OeRAD54 background: 11, clw; 12, cl37; 13, cl38; 14, San1ΔDUR3::Kan (chromosome VIII). n means phenotypically normal and * indicates sectored colony (see text for details). C) CHEF and Southern bots of the following samples (from left to right): 1, San1; in <i>rdh54Δ/rdh54Δ</i> background: 2, cl4; 3, cl7; 4, cl16; 5, cl23; 6, cl30 and 7, cl47. In CRAD54 background: 8, cl1 and 9, cl4; in OeRAD54 background: 10, clw; 11, cl37 and 12, cl38; 13, <i>top1Δ/top1Δ</i>cl18; 14, <i>xrs2Δ/xrs2Δ</i>cl6; 15, San1, hybridized with probes for <i>brx1, hal9</i> (upper line) and <i>cdc33</i> (not shown because identical to <i>hal9</i>) for chromosome XV and with <i>crp1</i>, <i>rim4</i> and <i>ste20</i> for chromosome VIII (bottom line). Two clones, obtained in <i>TOP1</i> and <i>XRS2</i> deletants, were also included in the CHEF (lanes 13 and 14); <i>brx</i> and <i>hal9</i> hybridization disclosed the complete loss of the native chromosome XV in <i>xrs2Δ/xrs2Δ</i>cl6 (lane 14).