Use of a ring chromosome and pulsed-field gels to study interhomolog recombination, double-strand DNA breaks and sister-chromatid exchange in yeast.
Use of a ring chromosome and pulsed-field gels to study interhomolog recombination, double-strand DNA breaks and sister-chromatid exchange in yeast.
复制标题
使用环状染色体和脉冲场凝胶研究酵母中的同源重组、双链 DNA 断裂和姐妹染色单体交换。
DOI：
10.1093/genetics/123.4.695
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发表时间：
1989
期刊：
影响因子：
3.3
通讯作者：
Mortimer,RK
中科院分区：
文献类型：
--
作者：
Game,JC;Sitney,KC;Cook,VE;Mortimer,RK
我们描述了一种使用脉冲场凝胶对酿酒酵母中的重组 DNA 分子、双链 DNA 断裂 (DSB) 和姐妹染色单体交换进行物理检测的系统。该系统利用了第三号染色体（Chr. III）的环状变体。该环染色体与线性同系物之间的减数分裂重组产生新分子，其大小在凝胶上可与任一亲本分子区分开来。我们证明这些重组分子也不存在于具有两个线性 Chr 的菌株中。 III 分子或 rad50 突变体，减数分裂重组有缺陷。结合分子终点，我们提供了减数分裂重组承诺时间的遗传评分数据。我们使用 X 射线来线性化圆形 Chr。 III，既开发一种测量 DSB 频率的灵敏方法，又作为检测部分源自姐妹染色单体交换的双倍大小圆的方法，我们发现这种交换在减数分裂期间很常见。
We describe a system that uses pulsed-field gels for the physical detection of recombinant DNA molecules, double-strand DNA breaks (DSB) and sister-chromatid exchange in the yeast Saccharomyces cerevisiae. The system makes use of a circular variant of chromosome III (Chr. III). Meiotic recombination between this ring chromosome and a linear homolog produces new molecules of sizes distinguishable on gels from either parental molecule. We demonstrate that these recombinant molecules are not present either in strains with two linear Chr. III molecules or in rad50 mutants, which are defective in meiotic recombination. In conjunction with the molecular endpoints, we present data on the timing of commitment to meiotic recombination scored genetically. We have used x-rays to linearize circular Chr. III, both to develop a sensitive method for measuring frequency of DSB and as a means of detecting double-sized circles originating in part from sister-chromatid exchange, which we find to be frequent during meiosis.