The use of microarrays in the analysis of gene expression is becoming widespread for many organisms including yeast. However, although a number of filamentous fungi have been fully or partially sequenced, microarray analysis is still in its infancy in these organisms. Here we describe the construction of microarrays for the fungus Aspergillus nidulans. A set of 4100 ESTs were isolated from conidial cDNA libraries and amplified by PCR. PCR products in 50% (v/v) DMSO were spotted onto Corning CMT-GAPS II slides using a Biorobotics Total Array System robot in 16 blocks of 17x16 spots 200mm apart. An experiment was designed to validate these arrays by monitoring the expression profile of known genes following the addition of 1% (w/v) glucose to wild-type A.nidulans cultures grown to mid-log phase in Vogel's minimal medium with ethanol as sole carbon source. Biomass samples for RNA extraction were taken from a 2 litre fermenter immediately before, (t=0) and at 1 hour, 2 hours and 4 hours after the addition of the glucose. RNA was extracted from biomass samples flash frozen in liquid nitrogen, labelled with cy3 or cy5 fluorescent dyes and hybridised to the arrays. Hybridisation profiles were analysed and quantified using Genepix software. The expression profiles following the glucose upshift will be presented and an assessment of the quality and reproducibility of the A.nidulans arrays discussed.
Fungal Genet. Newsl. 50 (Supl):abstract
Full conference title:
22nd Fungal Genetics Conference
- Fungal Genetics Conference 22nd (2001)