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首页> 外文期刊>Genomics >High-resolution fish on DNA fibers for low-copy repeats genome architecture studies
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High-resolution fish on DNA fibers for low-copy repeats genome architecture studies

机译：DNA纤维上的高分辨率鱼类可进行低拷贝重复基因组结构研究

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Low-copy repeats (LCRs) constitute 5% of the human genome. LCRs act as substrates for non-allelic homologous recombination (NAHR) leading to genomic structural variation. The aim of this study was to assess the potential of Fiber-FISH for LCRs direct visualization to support investigations of genome architecture within these challenging genomic regions. We describe a set of Fiber-FISH experiments designed for the study of the LCR22-2. This LCR is involved in recurrent reorganizations causing different genomic disorders. Four fosmid clones covering the entire length of the LCR22-2 and two single-copy BAC-clones, delimiting the LCR22-2 proximally and distally, were selected. The probes were hybridized in different multiple color combinations on DNA fibers from two karyotypically normal cell lines. We were able to identify three distinct structural haplotypes characterized by differences in copy-number and arrangement of the LCR22-2 genes and pseudogenes. Our results show that Multicolor Fiber-FISH is a viable methodological approach for the analysis of genome organization within complex LCR regions.
机译：低拷贝重复序列（LCR）占人类基因组的5％。 LCR作为非等位基因同源重组（NAHR）的底物，导致基因组结构变异。这项研究的目的是评估Fiber-FISH在LCR直接可视化方面的潜力，以支持在这些具有挑战性的基因组区域内进行基因组结构研究。我们描述了一组用于研究LCR22-2的Fiber-FISH实验。该LCR参与引起不同基因组疾病的复发性重组。选择了四个覆盖LCR22-2全长的fosmid克隆和两个单拷贝BAC克隆，从近端和远端划定了LCR22-2。将探针以不同的多种颜色组合在来自两种核型正常细胞系的DNA纤维上杂交。我们能够确定三种不同的结构单倍型，其特征在于LCR22-2基因和假基因的拷贝数和排列方式不同。我们的结果表明，Multicolor Fiber-FISH是分析复杂LCR区域内基因组组织的可行方法论方法。

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