Global genomic mapping, in Saccharomyces cerevisiae, of new suppressor alleles of the RNA polymerase I subunit rpa49 deletion.

ABSTRACT: In the yeast saccharoryces cerevisiae, RPA49 full-deletion mutants show a strong growth defect at 30°C and are unable to grow at 25°C. However, spontaneous suppressors that restore growth at 25°C are observed. We attend to identified new suppressor alleles after UV treatment. To mapped the genomic location of suppressor allele we have used a derivative of the so called \\"Genetic Interaction Mapping\\" (GIM) method (Decourty et al, 2008). Briefly, we have mated a yeast strain carrying the deletion of rpa49 and the a suppressor allele (query strain) with the pool of all haploid deletion mutants from the Euroscarf yeast gene deletion project. All deletions in strains from the Euroscarf collection are flanked by 2 unique 20 bases pair DNA tag (so-called Up-Tag and DN-tag) that allow detection on oligonucleotide micro-array. After sporulation in mass, spores carrying the rpa49 deletion, the suppressor allele and euroscarf deleted genes were selected and grow in a competition culture for 10 generations. As selected spores result from genetic re-association of allele from tested strains and euroscarf strain, all strains carrying a deleted genes from the euroscarf collection that is genetically link to the suppressor allele will be depleted in the culture. We next extract genomic DNA , amplified and labelled the tags. The relative quantity of each deletion tags in experiments with strain carrying suppressor alleles were estimated in a two color array experiment relative to the quantity of the tags in parallel experiments using two different wild-type strains.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Christophe Normand 

PROVIDER: E-MTAB-7831 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress