Abstract
To identify the genes encoding the proteins involved in cellulose degradation by the saprogenic basidiomycete Coprinus cinereus, differential display (DD) analysis based on Annealing Control Primer (ACP) system was carried out. Total RNAs were extracted from the mycelia of C. cinereus grown on the medium containing glucose or cellulose as the sole carbon source and then RT-PCR using ACP was performed in order to compare the amplification patterns of PCR products. As a result of DD analysis using 20 kinds of primer pairs, 10 kinds of cellulose culture-specific bands were detected. To confirm the transcript levels of the target genes, the specific primer to the nucleotide sequence of each band was prepared and RT-PCR analysis with the primers was carried out. The expression levels of the genes in the cellulose culture were higher than those in the glucose culture. In the database analysis using the deduced amino acid sequence, it was found that the genes encode various proteins such as not only the polysaccharide-degrading enzyme and the protein involved in carbohydrate metabolism but also cytochrome P450, retrotranspozon-relating protein and G protein-coupled receptor.
