Khademian Hossein, Giraut Laurène, Drouaud Jan, Mézard Christine
Institut National de la Recherche Agronomique (INRA), Centre de Versailles-Grignon, Institut Jean-Pierre Bourgin UMR1318 INRAAgroParisTech, Versailles Cedex, France.
Methods Mol Biol. 2013;990:177-90. doi: 10.1007/978-1-62703-333-6_18.
Meiotic recombination is essential for proper segregation of homologous chromosomes and thus for formation of viable gametes. Recombination generates either crossovers (COs), which are reciprocal exchanges between chromosome segments, or gene conversion not associated with crossovers (NCOs). Both kinds of events occur in narrow regions (less than 10 kb) called hotspots, which are distributed along chromosomes. While NCOs may represent a large fraction of meiotic recombination events in plants, as in many other higher eukaryotes, they have been poorly characterized due to the technical difficulty of detecting them. Here, we present a powerful approach, based on allele-specific PCR amplification of single molecules from pollen genomic DNA, allowing detection, quantification and characterization of NCO events arising at low frequencies at recombination hotspots.
减数分裂重组对于同源染色体的正确分离至关重要,因此对于形成有活力的配子也至关重要。重组产生交叉互换(COs),即染色体片段之间的相互交换,或者产生与交叉互换无关的基因转换(NCOs)。这两种事件都发生在称为热点的狭窄区域(小于10 kb),这些热点沿染色体分布。虽然在许多其他高等真核生物中,NCOs可能占植物减数分裂重组事件的很大一部分,但由于检测它们存在技术困难,它们的特征一直未得到充分研究。在这里,我们提出了一种强大的方法,基于从花粉基因组DNA中对单分子进行等位基因特异性PCR扩增,能够检测、定量和表征在重组热点以低频出现的NCO事件。