Bishop A J, Louis E J, Borts R H
Department of Cancer Cell Biology, Division of Molecular and Cellular Toxicology, Harvard School of Public Health, Boston, Massachusetts 02115, USA.
Genetics. 2000 Sep;156(1):7-20. doi: 10.1093/genetics/156.1.7.
Two yeast minisatellite alleles were cloned and inserted into a genetically defined interval in Saccharomyces cerevisiae. Analysis of flanking markers in combination with sequencing allowed the determination of the meiotic events that produced minisatellites with altered lengths. Tetrad analysis revealed that gene conversions, deletions, or complex combinations of both were involved in producing minisatellite variants. Similar changes were obtained following selection for nearby gene conversions or crossovers among random spores. The largest class of events involving the minisatellite was a 3:1 segregation of parental-size alleles, a class that would have been missed in all previous studies of minisatellites. Comparison of the sequences of the parental and novel alleles revealed that DNA must have been removed from the recipient array while a newly synthesized copy of donor array sequences was inserted. The length of inserted sequences did not appear to be constrained by the length of DNA that was removed. In cases where one or both sides of the insertion could be determined, the insertion endpoints were consistent with the suggestion that the event was mediated by alignment of homologous stretches of donor/recipient DNA.
克隆了两个酵母小卫星等位基因,并将其插入酿酒酵母的一个遗传定义区间。结合侧翼标记分析和测序,确定了产生长度改变的小卫星的减数分裂事件。四分体分析表明,基因转换、缺失或两者的复杂组合参与了小卫星变体的产生。在对随机孢子中的附近基因转换或交叉进行选择后,也获得了类似的变化。涉及小卫星的最大一类事件是亲本大小等位基因的3:1分离,这一类事件在以往所有小卫星研究中都会被遗漏。亲本和新等位基因序列的比较表明,在插入供体阵列序列的新合成拷贝时,受体阵列中的DNA一定被去除了。插入序列的长度似乎不受去除的DNA长度的限制。在可以确定插入一侧或两侧的情况下,插入端点与该事件由供体/受体DNA同源片段比对介导的推测一致。