Analysis of the Meiotic Transcriptome in Genetically Distinct Budding Yeasts Using High Density Oligonucleotide Arrays
Part of the Ernst Schering Research Foundation Workshop book series (SCHERING FOUND, volume 6)
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The last three years have witnessed a massive production of wholegenome expression data in the yeast field using two different types of gene arrays: commercially available high-density oligonucleotide arrays (GeneChips) and PCR-based gene arrays. In the case of GeneChips every yeast gene is represented by 20 oligonucleotides (each being a 25-mer) synthesized in situ onto a glass plate (which is then inserted into a cartridge for experimental manipulation; Fig. 1, panel b, top left image). Poly A+ RNA (e.g. prepared from cells at various stages of spore development; Fig. 1, panel a) is reverse transcribed into cDNA, labeled with a fluorophor and hybridized to the GeneChip. The fluorescence signal intensities of each set of 20 oligonucleotides are directly proportional to the mRNA concentration in the sample (Fig. 1, panel b, image of a GeneChip hybridization pattern). To eliminate the problem of cross hybridization a set of wild-type oligonucleotides (perfect match) are compared to a set of oligonucleotides containing a point mutation (mismatch) that destabilizes the DNA-DNA interaction. Two examples of a correct hybridization pattern (panel b, middle left) and a clear case of cross-hybridization (panel b, bottom left) are shown in Fig. 1.
KeywordsGene Array Fluorescence Signal Intensity Meiotic Gene Spore Development Sporulation Gene
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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- Chu S, Herskowitz I (1998) Gametogenesis in yeast is regulated by a transcriptional cascade dependent on Ndt80. Cell 1: 685 - 696Google Scholar
- Freire R, Murguia J, R, Tarsounas M, Lowndes N F, Moens PB, Jackson SP (1998) Human and mouse homologs of Schizosaccharomyces pombe radl+ and Saccharomyces cerevisiae RAD17: linkage to checkpoint control and mammalian meiosis. Genes & Dev 12: 2560 - 2573Google Scholar
- Gailus-Durner V, Xie J, Chintamaneni C, Vershon AK (1996) Participation of the yeast activator Abfl in meiosis-specific expression of the HOPI gene. Molecular & Cellular Biology 16: 2777 - 86Google Scholar
- Kupiec M, Byers B, Esposito R, Mitchell A (1997) Meiosis and Sporulation in Saccharomyces cerevisiae. Cold Spring Harbour Laboratory Press, 889 - 1036Google Scholar
- McKim K, Hayashi-Hagihara A (1998) mei-W68 in Drosophila melanogaster encodes a Spoil homolog: evidence that the mechanism for initiating meiotic recombination is conserved. Genes & Dev 12: 2932 - 2942Google Scholar
- Ozsarac N, Straffon MJ, Dalton HE, Dawes IW (1997) Regulation of gene expression during meiosis in Saccharomyces cerevisiae: SPR3 is controlled by both ABFI and a new sporulation control element. Molecular & Cellular Biology 17: 1152 - 9Google Scholar
- Phimister B (1999) Going global. Nature Genetics 21Google Scholar
- Ross-Macdonald P, Coelho P, Roemer T, Agarwal S, Kumar A, Jansen R, Cheung K, Sheehan A, Symoniatis D, Umansky L, Heidtman M, Nelson F, Iwasaki H, Hagers K, Gerstein M, Miller P, Roeder G, Snyder M (1999) Large-scale analysis of the yeast genome by transposon tagging and gene disruption. Nature 402: 413 - 418PubMedCrossRefGoogle Scholar
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