Freedman T, Pukkila P J
Department of Biology CB#3280, University of North Carolina at Chapel Hill 27599, USA.
Mol Gen Genet. 1997 Apr 28;254(4):372-8. doi: 10.1007/s004380050429.
We have used the polymerase chain reaction (PCR) method to monitor meiotic recombination in the basidiomycete Coprinus cinereus. We used DNA-mediated transformation to recover strains with modifications of the trp1 locus. The modifications were designed to introduce unique PCR priming sites separated by a homologous 2.4 kb region in which crossing over could occur. We showed that exchange occurred in this region at the frequency expected for a typical region of this genome (2.4 kb should correspond to a genetic length of 0.08 cM). We also detected products resulting from crossing over in DNAs extracted from cells in meiotic prophase. The assay should be useful for monitoring exchange in mutants that cannot complete meiosis.
我们已使用聚合酶链反应（PCR）方法监测担子菌灰盖鬼伞中的减数分裂重组。我们利用DNA介导的转化来获得trp1位点发生修饰的菌株。这些修饰旨在引入由一个同源的2.4 kb区域隔开的独特PCR引物位点，在该区域可能发生交叉互换。我们表明，在该区域发生了交换，其频率与该基因组典型区域预期的频率一致（2.4 kb应对应于0.08 cM的遗传长度）。我们还在减数分裂前期从细胞中提取的DNA中检测到了交叉互换产生的产物。该检测方法对于监测无法完成减数分裂的突变体中的交换应该是有用的。
