Cloning of USO1, yeast secretory gene, homologue from Aspergillus fungi

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In general, Aspergillus fungi secrete secrete large amount of protein in the medium but not yeast. We previously reported the isolation of the USO1 gene, which is required for intracellular protein transport. Uso1p is composed of globular head (780 aa), coiled-coil tail (1000 aa), and C-terminal acidic amino acids cluster.We searched EST database of Aspergillus nidulans and found a homologous sequence to the acidic domain of Uso1p and p115, mammalian homologue. We designed the primers based on the sequence and cloned a USO1 homologue gene from the genomic DNA of A.nidulans and named the gene as usoA.. We also isolated the corresponding cDNA and the sequence was analyzed. The usoA gene contains five introns and encodes a protein of 1103 amino acids. Predicted UsoA protein is composed of globular head (680 aa), coiled-coil tail (400 aa) and C-terminal acidic domain, which is conserved with Uso1p/p115. UsoA shows 46%, 29%, 29% and 26% identity to Uso1 or p115 from Neurospora crassa, S.cerevisiae, .5. pombe and bovine. We focus on investigation of intracellular protein transport in the fungus cells by repression of the usoA gene.