title: Reporter Gene Silencing Assays in Fission Yeast.
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.
authors.affiliation: Biology Department, Boston College, Chestnut Hill, Massachusetts 02467.
authors.forename: Hugh P
authors.initials: HP
authors.lastname: Cam
authors.affiliation: Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle NE2 4HH, United Kingdom.
authors.forename: Simon
authors.initials: S
authors.lastname: Whitehall
mesh_terms: D003470:Culture Media / Q000737:chemistry; D020868:Gene Silencing; D005800:Genes, Fungal; D017930:Genes, Reporter; D005827:Genetics, Microbial / Q000379:methods*; D006570:Heterochromatin / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods; D012568:Schizosaccharomyces / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D003470:Culture Media; D006570:Heterochromatin