Francis Kirk E, Lam Sandy Y, Harrison Benjamin D, Bey Alexandra L, Berchowitz Luke E, Copenhaver Gregory P
Department of Biology and Carolina Center for Genome Scientists, University of North Carolina, Chapel Hill, NC 27599, USA.
Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):3913-8. doi: 10.1073/pnas.0608936104. Epub 2007 Feb 23.
Recombination, in the form of cross-overs (COs) and gene conversion (GC), is a highly conserved feature of meiosis from fungi to mammals. Recombination helps ensure chromosome segregation and promotes allelic diversity. Lesions in the recombination machinery are often catastrophic for meiosis, resulting in sterility. We have developed a visual assay capable of detecting Cos and GCs and measuring CO interference in Arabidopsis thaliana. This flexible assay utilizes transgene constructs encoding pollen-expressed fluorescent proteins of three different colors in the qrt1 mutant background. By observing the segregation of the fluorescent alleles in 92,489 pollen tetrads, we demonstrate (i) a correlation between developmental position and CO frequency, (ii) a temperature dependence for CO frequency, (iii) the ability to detect meiotic GC events, and (iv) the ability to rapidly assess CO interference.
以交叉（COs）和基因转换（GC）形式存在的重组是从真菌到哺乳动物减数分裂的一个高度保守的特征。重组有助于确保染色体分离并促进等位基因多样性。重组机制中的损伤通常对减数分裂是灾难性的，会导致不育。我们开发了一种视觉检测方法，能够检测拟南芥中的COs和GCs，并测量CO干扰。这种灵活的检测方法利用在qrt1突变体背景下编码三种不同颜色花粉表达荧光蛋白的转基因构建体。通过观察92489个花粉四分体中荧光等位基因的分离，我们证明了（i）发育位置与CO频率之间的相关性，（ii）CO频率对温度的依赖性，（iii）检测减数分裂GC事件的能力，以及（iv）快速评估CO干扰的能力。
