Dataset Information


Profiling of Rgt1-regulated gene transcripts (37º C) in the fungal pathogen Candida albicans

ABSTRACT: C. albicans glucose sensing and signaling is important for full virulence in the mammalian host. To better understand this phenomenon, we investigated the Hgt4 glucose-sensing pathway, focusing on the downstream transcriptional repressor CaRgt1. CaRgt1-regulated gene transcripts were identified by comparing total RNA extracted from wild type and rgt1 mutant strains. Cells were grown at 37 degrees C in the presence of glycerol, conditions under which CaRgt1 was expected to repress gene transcription. Keywords: cell type comparison Overall design: A C. albicans genespecific microarray consisting of 70-mer oligonucleotides representing 6,346 of the 6,354 predicted ORFs in the annotated C. albicans genome assembly 19 was used. Each ORF was represented by a specific oligonucleotide, and one genome equivalent of this oligo was spotted three times per slide. To identify Rgt1-regulated transcripts, C. albicans cells from the wild type strain BWP17 or the rgt1 mutant strain CM18 were cultured in synthetic medium containing 5% glycerol at 37º C. Cells were harvested and RNA was extracted, labeled, and hybridized to the arrays. Each experiment consisted of a pair-wise competetive hybridization of total RNA samples derived from the wild type and mutant strains and included a reciprocal dye-flip replicate. Since two biological duplicates were tested for each strain, a total of four DNA microarray slides were used. Slides were scanned immediately after hybridization on a ScanArray express HT scanner (Perkin-Elmer). The scanner photomultiplier tube (PMT) values were set for optimal intensity with minimal background. An additional scan was done for each slide (low PMT) with the PMT such that <1% of the elements were saturated in order to characterise spots which were saturated at the higher PMT setting.

INSTRUMENT(S): WUCGS Candida albicans 6K v1.0

ORGANISM(S): Candida Albicans

SUBMITTER: Jessica Sexton 

PROVIDER: GSE7044 | GEO | 2007-03-01



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