Genome-wide mapping of transcription start sites in a ∆set2 strain
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ABSTRACT:
ORGANISM(S): Saccharomyces cerevisiae
PROVIDER: GSE62735 | GEO | 2015/01/14
SECONDARY ACCESSION(S): PRJNA265047
REPOSITORIES: GEO
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Project description:In cells lacking the histone methyltransferase Set2, initiation of RNA polymerase II transcription occurs inappropriately within the protein-coding regions of genes, rather than being restricted to the proximal promoter. Here, we mapped the transcripts produced in an S. cerevisiae strain lacking Set2, and applied rigorous statistical methods to identify sites of cryptic transcription at high resolution. Wild type (BY4741) and set2â (BY4741) strains were grown at 30°C in YPD (1% yeast extract, 2% peptone, 2% dextrose) to an OD600 of 0.6-0.8. For each of the three replicates, Total RNA was extracted by acid-phenol method (Xiao et al. 2003). Double-stranded cDNA was prepared using an Invitrogen SuperScript⢠(Cat No. 11917-010) primed with Oligo(dt) and random hexamers. For each replicate, the wt and set2â cDNA were independetly fluorescently labeled and comparatively hybridized to high-resolution 385K Saccharomyces cerevisiae CGH arrays (2005-08-16_SCER_WG_CGH) with Tm-normalized probes. In one of the replicates, assignment of the fluorescent label was reversed.
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Project description:The goals of this study are to compare different gene expressions for Penicillium oxalicum wild type strain (WT), and set2 knockout strain (Δset2) in different carbon sources. The deletion of set2 upregulated genes involved in oxidation- reduction process, extracellular region, and plasma membrane ATP synthesis coupled proton transport both with 2% glucose or 1% cellulose and 1% wheat bran as carbon sources. We find the expression levels of 20 secondary metabolism gene clusters were upregulated or downregulated under different carbon sources in Δset2. This study provides the information that SET2 function are required in conidiation and hydrolase activity of P. oxalicum.
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