Abstract
DNA within the yeast nucleus is spatially organized. Yeast telomeres cluster together at the nuclear periphery, centromeres cluster together near the spindle pole body, and both the rDNA repeats and tRNA genes cluster within the nucleolus. Furthermore, the localization of individual genes to subnuclear compartments can change with changes in transcriptional status. As such, yeast researchers interested in understanding nuclear events may need to determine the subnuclear localization of parts of the genome. This chapter describes a straightforward quantitative approach using immunofluorescence and confocal microscopy to localize chromosomal loci with respect to well characterized nuclear landmarks.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 569-580 |
| Number of pages | 12 |
| Journal | Methods in enzymology |
| Volume | 470 |
| Issue number | C |
| DOIs | |
| State | Published - 2010 |
Funding
The authors thank John Sedat for sharing the GFP-Lac repressor and the LacO array plasmids, Francoise Stutz for the GAL2 LacO plasmid, Kevin Redding for developing the immunofluorescence protocol on which these methods are based and members of the Brickner laboratory for helpful comments on this chapter. J. H. B. is supported by NIH grant GM080484, the W. M. Keck Foundation and the Baldwin Fund for Biomedical Research.
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology