Project description:In this study, we constructed three isogenic strains of S96 yrr1Δ background (its native YRR1 gene was knocked out) carrying three different YRR1 alleles, YRR1_S96, YRR1_YJM789, YRR1_S96-I775E, respectively. We then conducted chromatin immuno-precipitation followed by high-throughput sequencing (ChIP-Seq) for Yrr1 protein on the three strains grown in Yeast Peptone Dextrose medium (YPD) and YPD + 4NQO.
Project description:We measured the response of S. cerevisiae to arrest in the presence of alpha factor. These were collected in support of a related DNaseI-sequencing study. Keywords: Alpha-factor arrest S.cerevisiae R276 (MATa ura3Δ0 leu2Δ0 his3Δ1 met15Δ0 bar1Δ::KanMX) (C. Boone, University of Toronto; S288c background derived from BY4741), was cultured overnight in 50 ml rich medium (YPD) at 30°C, diluted into 500 ml fresh YPD to an OD660 of ~0.8, and treated with yeast α-factor (Sigma-Aldrich) at a final concentration of 50 ng / ml. This culture was incubated at 30°C with shaking for 3 hours (final OD660 ~1). After this treatment, approximately 90% of the cells had formed mating projections when checked by light microscopy. Total RNA from these cells was isolated using hot acidic phenol. 50 μg of total RNA was treated with Turbo Dnase (Ambion), and checked for integrity using a Bioanalyzer 2100 (Agilent). Total RNA was labeled according to the manufacturer’s protocol and applied to Affymetrix Yeast 2.0 arrays. Data were analyzed using the “affy” package from Bioconductor.
Project description:We performed massive screening of the genes in yeast that were involved in the tolerance to isopropanol using the non-essential genes deleted yeast collection, and identified sixty-five disruptants that grew slower than the wild type strain in the presence of isopropanol. The isopropanol sensitive mutants were tested to know their behaviour under other alcohol stresses. Besides, we conducted microarray analysis to reveal the transcriptional response to isopropanol stress in yeast. Our results certainly provide new insights into yeast response to C3 alcohol isopropanol. Strain BY4743 was used in this test. The prepared cells were inoculated into 120 ml of fresh media (YPD without isopropanol, and YPD with 20 g/l or 40 g/l of isopropanol) in 500-ml Sakaguchi-flasks. They were cultivated for 3 h at 30M-BM-0C with 130-rpm shaking. At least three independent experiments were performed at each concentration of isopropanol.
Project description:In this study, we constructed three isogenic strains of S96 yrr1Δ background (its native YRR1 gene was knocked out) carrying three different YRR1 alleles, YRR1_S96, YRR1_YJM789 and YRR1_S96-I775E, respectively. We then conducted RNA deep sequencing (RNA-Seq) on the three strains grown in Yeast Peptone Dextrose medium (YPD), YPD + 4NQO and Yeast Peptone glycerol medium (YPglycerol).
