Kim K-D, Iwasaki O, Noma K
The Wistar Institute, Philadelphia, PA, United States.
The Wistar Institute, Philadelphia, PA, United States.
Methods Enzymol. 2016;574:167-180. doi: 10.1016/bs.mie.2016.04.003. Epub 2016 Jun 1.
Recent genomic studies have revealed that chromosomal structures are formed by a hierarchy of organizing processes ranging from gene associations, including interactions among enhancers and promoters, to topologically associating domain formations. Gene associations identified by these studies can be characterized by microscopic analyses. Fission yeast is a model organism, in which gene associations have been broadly mapped across the genome, although many of those associations have not been further examined by cell biological approaches. To address the technically challenging process of the visualization of associating gene loci in the fission yeast nuclei, we provide, in detail, an IF-FISH procedure that allows for covisualizing both gene loci and nuclear structural markers such as the nuclear membrane and nucleolus.
最近的基因组研究表明，染色体结构是由一系列组织过程形成的层次结构，从基因关联（包括增强子和启动子之间的相互作用）到拓扑相关结构域的形成。这些研究确定的基因关联可以通过显微镜分析来表征。裂殖酵母是一种模式生物，其中基因关联已在全基因组范围内广泛绘制，尽管其中许多关联尚未通过细胞生物学方法进一步研究。为了解决在裂殖酵母细胞核中可视化关联基因位点这一技术上具有挑战性的过程，我们详细提供了一种间接免疫荧光-荧光原位杂交（IF-FISH）程序，该程序允许同时可视化基因位点和核结构标记物，如核膜和核仁。
