Expression data from sexual flocculation cells of S. pombe
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ABSTRACT: The h+ and h- haploid cells of S. pombe mixtures flocculated and formed diploid cells when cultured in carbon limited YEPME medium. We first cultured S. pombe h+ YHL6381 and h- SPQ-01 cells respectively in YEPD medium and then mixed them at an 1:1 ratio and further cultured in YEPME medium. Sexual flocculation occured. We used microarrays to detail the the global programme of gene expression of flocculated YHL6381 and SPQ-01 mixtures cultured in YEPME, compared with the non flocculated cells mixtures cultured in YEPD, and identified distinct classes of up-regulated genes during this process. We first cultured S. pombe h+ YHL6381 and h- SPQ-01 cells respectively in YEPD medium and then mixed them at an 1:1 ratio and further cultured in a new medium for 90min. The flocculated cells mixtures cultured in YEPME named YSD, while the non flocculated cells mixtures cultured in YEPD named YSM.
Project description:The h+ and h- haploid cells of S. pombe mixtures flocculated and formed diploid cells when cultured in carbon limited YEPME medium. We first cultured S. pombe h+ YHL6381 and h- SPQ-01 cells respectively in YEPD medium and then mixed them at an 1:1 ratio and further cultured in YEPME medium. Sexual flocculation occured. We used microarrays to detail the the global programme of gene expression of flocculated YHL6381 and SPQ-01 mixtures cultured in YEPME, compared with the non flocculated cells mixtures cultured in YEPD, and identified distinct classes of up-regulated genes during this process.
Project description:Deleting ribosomal protein genes caused haploid cells of S. pombe flocculated in full nutrition YEPD medium. We deleted rpl32-1 and rpl32-2 gene respectively and these two mutant cells cultured in YEPD medium flocculated. We used microarrays to detail the global programme of gene expression between non flocculated rpl32-1 deleted and rpl32-2 deleted, compared with the non flocculated cells SPQ-01, and identified distinct classes of up-regulated genes during this process. Yeast cells cultured in YEPD medium in the log phase were selected for RNA extraction and hybridization on Affymetrix microarrays. The rpl32-1 deleted cells were conformed as SD1 and rpl32-2 deleted cells were conformed as SD2. Control cells SPQ-01 were conformed as WT.
Project description:Deleting ribosomal protein genes caused haploid cells of S. pombe flocculated in full nutrition YEPD medium. We deleted rpl32-1 and rpl32-2 gene respectively and these two mutant cells cultured in YEPD medium flocculated. We used microarrays to detail the global programme of gene expression between non flocculated rpl32-1 deleted and rpl32-2 deleted, compared with the non flocculated cells SPQ-01, and identified distinct classes of up-regulated genes during this process.
Project description:Transcription profiles across the cell cycle of yox1 deleted yeast cells. BY2399 cells (yox1 delete cells isogenic with W303a) were arrested with alpha factor in YEPD media. After release from arrest cells were sampled every 5 min for 2 hr. Keywords: cell cycle time course Cells were synchronized with alpha factor and sampled every 5 min across 2 cell cycles. A total of 25 samples were analyzed. Technical replicates were included. cDNA of the cell cycle samples were labelled with Cy5. For Cy3 labelling, asynchronous yeast population was used.
Project description:ChIP-chip to determine the regulation of the K. lactis hsgs (ChIP of MATa1, MATalpha2, and RME1). The MATa1 and MATalpha2 ChIPs were performed in an a/alpha cell using N-terminally HA-tagged proteins and the RME1 ChIPs were perfomed in an a cell using C-terminally myc-tagged protein. For the RME1 ChIPs, the cells grown with out phosphate. For the MATa1 andMATalpha2 ChIPs the cells were grown in YEPD. Epitope tagged strains were compared to untagged control strains. Two biological replicates were preformed for the RME1 ChIP. For the MATa1 and MATalpha2 ChIP, peaks were considered indicative of binding if both MATa1 and MATalpha2 showed enrichment above the untagged control.
Project description:Transcriptional profiling was performed on cells grown in YEPD medium at 28 M-0C to an OD600 nm = 1.8, in case of cells grown in exponential phase; alternatively, cells were growth in 1% N-acetylglucosamine and 50 mM citrate buffer pH6.0 at 28 M-0C for different times points (15, 60 and 180 minutes) to induce yeast-hypha transition.
Project description:Goal was to identify yeast genes whose expression changed as a function of the shift from growth in bulk culture to growth in an air-liquid interfacial biofilm. Experiment Overall Design: Cells were grown 24 h at 30 C in YEPD to a density of about 500,000,000 cells/ml. At harvest, sugar was found to be depleted as measured by an enzymatic dip stick (Diastic, Bayer). Cells were pelleted and washed twice in sterile distilled water by centrifugation and then diluted 10-fold into 100 ml of Flor medium (YNB + 4% ethanol + leucine + histidine + uracil) in triplicate 250 ml beakers. Cultures were then grown statically in Flor medium at 27 C. Within a few hours following inoculation, the bulk liquid appeared to be clear, no biofilm was evident by visual observation, and a layer of settled cells was evident at the bottom of the beakers. After 48 h, a visible air-liquid interfacial biofilm covered the entire surface while the thickness of the layer of settled cells appeared unchanged. After 48 h, biofilm cells (floaters) were collected by aseptic aspiration. Once the biofilm cells were removed, cells at the bottom of the beaker (sinkers) were collected similarly. Cells from both populations were washed once in sterile distilled water by centrifugation prior to RNA isolation. Significant cell clumping was evident in both populations of cells by microscopic observation. While cell viability was estimated by plating on YEPD, the resultant cfu/ml values could not be directly correlated with cell counts in a hemacytometer because clumps of cells containing at least one viable cell presumably produced only a single colony. Further, counting individual cells accurately in the numerous clumps containing large numbers of cells was not possible. Nonetheless, when cells were counted (clumps were counted as single cells) and compared to cfu/ml for the same suspension, the cfu/ml values were about 2-fold higher than the corresponding values for cell counts using the hemacytometer for both biofilm and bottom layer cells.
Project description:PDR mutants , strain YYA100 (pdr1?::KanMx6, pdr3?::His3Mx6) from Mol Biol Cell. 2007 December; 18(12): 49324944, was grown to OD 1-1.5 in YEPD and hybridyzed to an expression array in one channel with a wild-type strain reference RNA on the other channel