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首页> 外文期刊>Molecular biology of the cell >Long-range interphase chromosome organization in Drosophila: A study using color barcoded fluorescence in situ hybridization and structural clustering analysis
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Long-range interphase chromosome organization in Drosophila: A study using color barcoded fluorescence in situ hybridization and structural clustering analysis

机译：果蝇的远期相间染色体组织：使用彩色条形码荧光原位杂交和结构聚类分析的研究

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摘要

We have developed a color barcode labeling strategy for use with fluorescence in situ hybridization that enables the discrimination of multiple, identically labeled loci. Barcode labeling of chromosomes provides long-range path information and allows structural analysis at a scale and resolution beyond what was previously possible. Here, we demonstrate the use of a three-color, 13-probe barcode for the structural analysis of Drosophila chromosome 21, in blastoderm stage embryos. We observe the chromosome to be strongly polarized in the Rab1 orientation and for some loci to assume defined positions relative to the nuclear envelope. Our analysis indicates packing similar to15- to 28-fold above the 30-nm fiber, which varies along the chromosome in a pattern conserved across embryos. Using a clustering implementation based on rigid body alignment, our analysis suggests that structures within each embryo represent a single population and are effectively modeled as oriented random coils confined within nuclear boundaries. We also found an increased similarity between homologous chromosomes that have begun to pair. Chromosomes in embryos at equivalent developmental stages were found to share structural features and nuclear localization, although size-related differences that correlate with the cell cycle also were observed. The methodology and tools we describe provide a direct means for identifying developmental and cell type-specific features of higher order chromosome and nuclear organization.
机译：我们已经开发了一种用于荧光原位杂交的彩色条形码标记策略，该策略可以区分多个相同标记的基因座。染色体的条形码标记可提供远距离路径信息，并允许以超出以前可能的规模和分辨率进行结构分析。在这里，我们展示了在胚盘期胚胎中使用三色，13探针条形码对果蝇21号染色体进行结构分析。我们观察到该染色体在Rab1方向上强烈极化，并且某些基因座相对于核包膜采取了确定的位置。我们的分析表明，在30 nm光纤上方堆积大约15到28倍，堆积情况沿染色体变化，在整个胚胎中均保持不变。使用基于刚体对准的聚类实现，我们的分析表明，每个胚胎内的结构代表单个种群，并被有效地建模为局限于核边界内的定向随机线圈。我们还发现已经开始配对的同源染色体之间的相似性增加。尽管也观察到与细胞周期相关的大小相关差异，但处于同等发育阶段的胚胎中的染色体具有相同的结构特征和核定位。我们描述的方法和工具为鉴定高阶染色体和核组织的发育和特定细胞类型的特征提供了直接手段。

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