Conservation of Meiotic Recombination Sites in the Human Genome

描述（申请人提供）：减数分裂重组是重要的生物学过程之一，可能导致许多人类疾病。在人类中，已知减数分裂重组主要聚集在称为重组热点的某些“重组位点”。虽然重组热点经常在不同人类个体的相同位置发现，但对它们的保守性知之甚少。在拟议的研究中，假设人类基因组中的重组热点是高度保守的，但会发生重大变化，这可能是产生新位点的原因，并削弱和消除现有位点。为此，将使用18号染色体的短臂作为研究的受试者。在该项目的第一步中，来自三名人类男性的9，000个单精子样本，每个3，000个，将在全基因组扩增（WGA）后在4，000个单核苷酸多态性（SNP）位点进行基因型分析。这将通过使用P.I.开发的高通量基因分型方法来完成。的实验室。完成这一轮分析后，将检测重组热点，并将其定位于染色体臂沿着长的~4.8 kb区域。通过对位于包含交叉区域的另外4，000个SNP进行基因分型，将这些点进一步定位于<1 kb区域。将对所有包含交叉的区间（n = 500 - 600）进行保守性分析。将通过使用先前报道的方法分析合并的精子样本，确定来自两个种族的20名精子供体在所有这些时间间隔内的重组率，该方法允许选择性扩增和检测含有重组交叉的序列。由此产生的数据可能首次用于系统地研究大规模重组热点的保守性，这将有助于了解人类基因组遗传结构的动态和设计疾病关联研究。

DESCRIPTION (provided by applicant): Meiotic recombination is one of the important biological processes, and may be responsible for many human diseases. In humans, meiotic recombination is known to be clustered mostly at certain "recombination sites" known as recombination hotspots. Although recombination hotspots are often found at the same locations in different human individuals, very little is known about their conservation. In the proposed study, a hypothesis that recombination hotspots in the human genome are highly conserved but are subjected to substantial changes which may be responsible for the generation of new sites, and weakening and extinguishing existing sites. To do so, the short arm of chromosome 18 will be used as the subject for the study. In the first step of the project, 9,000 single sperm samples from three human males, 3,000 each, will be subjected to genotype analysis at 4,000 loci of Single Nucleotide Polymorphisms (SNPs) after whole genome amplification (WGA). This will be done by using a high-throughput genotyping approach developed in the P.I.'s laboratory. Completion of this round of analysis will detect and localize the recombination hotspots to ~4.8 kb regions along length of the chromosomal arm. These spots will be further localized to <1 kb regions by genotyping an additional 4,000 SNPs located in the regions containing crossovers. All crossover-containing intervals (n = 500 to 600) will be subjected to the conservation analysis. Recombination rates in all these intervals will be determined in 20 sperm donors from two ethnic groups by analyzing pooled sperm samples using a previously reported method allowing selectively amplification and detection of sequences containing recombination crossovers. The resulting data may be used, for the first time, to systematically examine the conservation of recombination hotspots on a large-scale, which will help understand the dynamics of the genetic structure of the human genome and design disease association studies.