Measurement of spatial proximity and accessibility of chromosomal loci in Saccharomyces cerevisiae using Cre/loxP site-specific recombination.
Measurement of spatial proximity and accessibility of chromosomal loci in Saccharomyces cerevisiae using Cre/loxP site-specific recombination.
复制标题
使用 Cre/loxP 位点特异性重组测量酿酒酵母中染色体位点的空间邻近性和可达性。
DOI：
10.1007/978-1-59745-527-5_5
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Burgess,SeanM
中科院分区：
文献类型：
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作者：
Lui,Doris;Burgess,SeanM
已经开发了几种方法来测量芽殖酵母减数分裂过程中同源染色体之间的相互作用。其中包括使用荧光原位杂交 (FISH) 对固定扩散核进行细胞学分析 (1, 2)、活细胞中 GFP 标记的染色体位点可视化 (3) 以及染色体构象捕获 (3C)(4)。在这里，我们描述了一种定量遗传测定，该测定使用外源位点特异性重组来监测芽殖酵母活细胞中两个确定基因座之间的同源关联水平(5)。我们使用 Cre/loxPassay 对芽殖酵母中的核结构和减数分裂同源配对进行了基因剖析。从该测定中获得的数据报告了位于同一菌株内的两个染色体基因座的相对空间接近性或可及性，并且可以与不同突变菌株的测量结果进行比较。
Several methods have been developed to measure interactions between homologous chromosomes during meiosis in budding yeast. These include cytological analysis of fixed, spread nuclei using fluorescence in situ hybridization (FISH)(1, 2), visualization of GFP-labeled chromosomal loci in living cells(3), and Chromosome-Conformation Capture (3C)(4). Here we describe a quantitative genetic assay that uses exogenous site-specific recombination to monitor the level of homolog associations between two defined loci in living cells of budding yeast(5). We have used the Cre/loxPassay to genetically dissect nuclear architecture and meiotic homolog pairing in budding yeast. Data obtained from this assay report on the relative spatial proximity or accessibility of two chromosomal loci located within the same strain and can be compared to measurements from different mutated strains.