Unknown,Transcriptomics,Genomics,Proteomics

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Expression data from sexual flocculation cells of S. pombe

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.

ORGANISM(S): Schizosaccharomyces pombe

SUBMITTER: Rongpeng Li

PROVIDER: E-GEOD-43250 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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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.

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Expression data from sexual flocculation cells of S. pombe

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