Reporter Gene Silencing Assays in Fission Yeast
- 1Biology Department, Boston College, Chestnut Hill, Massachusetts 02467;
- 2Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle NE2 4HH, United Kingdom
Abstract
Reporter gene silencing assays provide a facile method for assessing the function of heterochromatin in Schizosaccharomyces pombe. They use strains containing auxotrophic markers (commonly ura4+ or ade6+) located within a heterochromatic region. Transcriptional silencing of these reporters can be assessed by plating serial dilutions of cells onto minimal agar. In addition, silencing of ura4+ renders cells resistant to 5-fluoroorotic acid (5-FOA) and ade6+ silencing results in red colony color on adenine-limiting agar. Various reporters for each of the major heterochromatic domains (telomeres, centromeres, and the mating type locus) are available and, importantly, transcriptional silencing is correlated with the proper function of these regions.
Footnotes
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↵3 Correspondence: hugh.cam{at}bc.edu; simon.whitehall{at}newcastle.ac.uk
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From the Fission Yeast collection, edited by Iain M. Hagan, Antony M. Carr, Agnes Grallert, and Paul Nurse.
- © 2016 Cold Spring Harbor Laboratory Press
