Abstract:
During meiosis, homologous chromosomes reciprocally exchange segments of DNA via the formation of crossovers. However, the frequency and position of crossover events along chromosomes are not random. Each chromosome must receive at least one crossover, and the formation of a crossover at one location inhibits the formation of additional crossovers nearby. These crossover patterning phenomena are referred to as \"crossover assurance\" and \"crossover interference,\" respectively. One key method for quantifying meiotic crossover patterning is to immunocytologically measure the position and intensity of crossover-associated protein foci along the length of meiotic prophase I chromosomes. This approach was recently used to map the position of a conserved E3 ligase, HEI10, along Arabidopsis pachytene chromosomes, providing experimental support for a novel mechanistic \"coarsening model\" for crossover patterning. Here we describe a user-friendly method for automatically measuring the position and intensity of recombination-associated foci along meiotic prophase I chromosomes that is broadly applicable to studies in different eukaryotic species.
摘要:
在减数分裂期间,同源染色体通过交叉的形成相互交换DNA片段。然而,沿着染色体的交叉事件的频率和位置不是随机的。每个染色体必须至少接受一次交叉,并且在一个位置形成交叉会抑制附近形成额外的交叉。这些交叉图案现象被称为“交叉保证”和“交叉干扰”,\"分别。定量减数分裂交换模式的一个关键方法是免疫细胞学测量沿着减数分裂前期I染色体长度的交换相关蛋白灶的位置和强度。这种方法最近被用来绘制保守的E3连接酶的位置,HEI10,沿着拟南芥粗线质染色体,为交叉图案化的新型机械“粗化模型”提供实验支持。在这里,我们描述了一种用户友好的方法,用于自动测量减数分裂前期I染色体上重组相关灶的位置和强度,该方法广泛适用于不同真核生物物种的研究。