Project description:This research work investigates the expression of the genes involved in flavor compound production in two hybrids between Saccharomyces cerevisiae and S. kudriavzevii under low (12°C) and moderate fermentation temperatures (28°C).
Project description:Gene expression of different flocculent (HS 10 and HS 34) and powdery (HS 12 and HS 37) yeast strains compared to each other during exponential and stationary growth phase was analysed. The isolation of RNA was done by disruption of the cells under liquid nitrogen using mortar and pistil and then the Qiagen RNeasy Midi Kit with some modifications within the manufacturers´ protocol and the Qiagen RNase-Free DNase Set were applied. 6 µg of the total RNA per sample was used for each microarray experiments. The indirect labelling by the tyramide-signal-amplification method (MicromaxTM TSATM labelling and detection Kit from Perkin Elmer life sciences) was used to increase the Cy3 and Cy5 signals of microarray detection. Each cDNA containing Biotin- and Fluorescein-nucleotides respectively was purified with a QIAquick PCR purification kit and suspended in 11 µl of the formamide containing hybridization buffer. The slides were hybridized at 42°C over night under a cover slip. The microarrays were scanned by the Axon 4000B scanner; image intensity data were extracted and analysed with GenePix® Pro 6.0 software. Data from different scans of Dye-swap experiment were extracted by GenePix Pro 6.0 software, normalized and united. An outliertest has been applied in order to find outliers amongst the gene replicats. Subsequently, a t-test (1% and 5% probability of error) has been used in order to find regulated genes. Keywords: sorted yeast cells
Project description:Saccharomyces cerevisiae is an excellent microorganism for industrial succinic acid production, but high succinic acid concentration will inhibit the growth of Saccharomyces cerevisiae then reduce the production of succinic acid. Through analysis the transcriptomic data of Saccharomyces cerevisiae with different genetic backgrounds under different succinic acid stress, we hope to find the response mechanism of Saccharomyces cerevisiae to succinic acid.
Project description:Intact nuclei from an asynchronous population of W303 Saccharomyces cerevisiae in log-phase growth were subjected to a 16-minute DNase I digestion (0.1 U/μL) at 37 °C. DNA was then recovered, and single-end Illumina sequencing libraries were prepared using the Crawford DNase-seq method (Song and Crawford, 2010).
Project description:Comparison of the transcriptomes of Saccharomyces cerevisiae wild type FY23 and a PDE2 deletion mutant DJ28. Keywords = PDE2 Keywords = Ras/cAMP pathway Keywords: other
Project description:In this project, we follow the temporal proteome dynamics during heat stress response in a model organism Saccharomyces cerevisiae. Using label-free quantification we determine changes in the yeast proteome at different times after mild temperature shift, from 30˚C to 37˚C. The time points measured were 0 min (30˚C, no stress) and then at 10 min, 30 min, 60 min, 120 min and 240 min after cells were transferred to 37 °C. For each time point, four biological replicates were collected for the haploid wild type (BY4742, Mat ALPHA, his3Δ1; leu2Δ0; lys2Δ0; ura3Δ0; YJL088w::kanMX4) strain and a haploid strain with a chaperone Ssb1p deletion (ssb1Δ).
Project description:Gene expression of different flocculent (HS 10 and HS 34) and powdery (HS 12 and HS 37) yeast strains compared to each other during exponential and stationary growth phase was analysed. The isolation of RNA was done by disruption of the cells under liquid nitrogen using mortar and pistil and then the Qiagen RNeasy Midi Kit with some modifications within the manufacturers´ protocol and the Qiagen RNase-Free DNase Set were applied. 6 µg of the total RNA per sample was used for each microarray experiments. The indirect labelling by the tyramide-signal-amplification method (MicromaxTM TSATM labelling and detection Kit from Perkin Elmer life sciences) was used to increase the Cy3 and Cy5 signals of microarray detection. Each cDNA containing Biotin- and Fluorescein-nucleotides respectively was purified with a QIAquick PCR purification kit and suspended in 11 µl of the formamide containing hybridization buffer. The slides were hybridized at 42°C over night under a cover slip. The microarrays were scanned by the Axon 4000B scanner; image intensity data were extracted and analysed with GenePix® Pro 6.0 software. Data from different scans of Dye-swap experiment were extracted by GenePix Pro 6.0 software, normalized and united. An outliertest has been applied in order to find outliers amongst the gene replicats. Subsequently, a t-test (1% and 5% probability of error) has been used in order to find regulated genes. Keywords: sorted yeast cells Microarray experiments were realised with dye swap.
Project description:A total of 12 WW domains from Saccharomyces cerevisiae were expressed and purified as fusion proteins to either GST or MBP. The fusion proteins were chemically biotinylated and applied to duplicate protein microarrays. Data processing revealed a total of 587 interactions between the domains and 207 proteins. Most of these interactions have not been previously observed. Keywords: Protein microarray analysis of yeast WW domains
Project description:To gather more in-depth knowledge of the Mtl1p mechanosensor's role in Saccharomyces cerevisiae metabolism, we conducted a comparative metabolomic analysis of two Saccharomyces cerevisiae strains: the wild type and mtl1Δ, which carries a deletion of the mechanosensor Mtl1p. Both strains were grown under normal conditions at 27°C. The most significant metabolic changes between these strains were related to amino acid metabolism, purine metabolism, and carboxylic acid metabolism.
Project description:To gather more in-depth knowledge of the Mtl1p mechanosensor's role in Saccharomyces cerevisiae metabolism, we conducted a comparative metabolomic analysis of two Saccharomyces cerevisiae strains: the wild type and mtl1Δ, which carries a deletion of the mechanosensor Mtl1p. Both strains were grown under normal conditions at 27°C. The most significant metabolic changes between these strains were related to amino acid metabolism, purine metabolism, and carboxylic acid metabolism.
