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Mer1D_SK1, mer2D_SK1, mer3D_SK1, spo22D_SK1, and spo70D_SK1 on expression/splicing microarrays

ABSTRACT: mer1D_SK1, mer2D_SK1, mer3D_SK1, spo22D_SK1, spo70D_SK1 late 9h meiotic time point. mer1D, mer2D, mer3D, spo22D diploid, and spo70D diploid was generated in the SK1 strain background (ATCC: MYA-2089) by PCR-mediated cassette-based gene replacement where the MER1, MER2, MER3, SPO22, or SPO70 ORF respectively was replaced with HIS3 from S. kluyveri then the transformants were sporulated and the homozygous deletion was verified by PCR.

ORGANISM(S): Saccharomyces cerevisiae

PROVIDER: GSE24678 | GEO | 2011/04/05

SECONDARY ACCESSION(S): PRJNA133623

REPOSITORIES: GEO

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mer1D_SK1, mer2D_SK1, mer3D_SK1, spo22D_SK1, and spo70D_SK1 on expression/splicing microarrays

Project description:mer1D_SK1, mer2D_SK1, mer3D_SK1, spo22D_SK1, spo70D_SK1 late 9h meiotic time point. mer1D, mer2D, mer3D, spo22D diploid, and spo70D diploid was generated in the SK1 strain background (ATCC: MYA-2089) by PCR-mediated cassette-based gene replacement where the MER1, MER2, MER3, SPO22, or SPO70 ORF respectively was replaced with HIS3 from S. kluyveri then the transformants were sporulated and the homozygous deletion was verified by PCR. Meiotic time point =9h; Dye swap; Reference WT_SK1 9h

2011-04-05 | E-GEOD-24678 | biostudies-arrayexpress

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Project description:Synchronized mer1D_SK1 meiotic time course. mer1D diploid was generated in the SK1 strain background (ATCC: MYA-2089) by PCR-mediated cassette-based gene replacement where the MER1 ORF was replaced with HIS3 from S. kluyveri then the transformants were sporulated and the homozygous deletion was verified by PCR.

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Project description:Replicate cultres of Saccharomyces cerevisiae strain VL3 (a commercial wine strain) were inititaed with and without the the co-inoculation of a second yeast species, Pichia kluyveri (PKKR1). In addition replicate cultures of Pichia kluyveri were initiated. The media for all cultures was sterile Sauvignon blanc grape juice. Triplicates of each treatment were destructively harvested at days 2, 9 and 16 of fermentation and RNA extracted. RNA from S. cerevisiae could not be preferentially extracted from co-ferments. P. kluyveri genome has not been sequenced and so homology to S. cerevisiae probes on the array could not be accurately estimated. We extracted RNA from the P. kluyveri single ferments and hybridised them to the S. cerevisiae arrays to control for this. These species diverged over 100 MYA, and we did not observe abundant homology by way of cross hybridisation. The mean probe intensity for P. kluyveri cDNA was 17-fold lower than for S. cerevisiae, but some probes reported reasonable signal. The intensity distribution of these probes fit a log-normal distribution (P<0.01). 97% of the distribution fell below one-thirtieth of the S. cerevisiae maximum intensity: the remaining 3%, comprising 429 probes, potentially have reasonable homology to P. kluyveri cDNA and we removed these from all analyses. Please see associated additional file 'removePKKR1.r' R code to achieve this.

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