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

一般情况下，曲霉在培养基中分泌大量蛋白质，而不分泌酵母。我们之前曾报道过USO1基因的分离，这是细胞内蛋白质运输所必需的。Uso1p由球形头部(780aa)、盘绕尾部(1000aa)和C-末端酸性氨基酸簇组成，搜索Nidulans的EST数据库，发现了与哺乳动物同源物Uso1p和p115的酸性结构域的同源序列。根据该序列设计了相应的引物，克隆了一个USO1同源基因，命名为usoA。我们还分离了相应的基因，并对其进行了序列分析。UsoA基因包含5个内含子，编码1103个氨基酸的蛋白质。预测的UsoA蛋白由球形头部(680aa)、盘绕尾部(400aa)和C-末端酸性结构域组成，与Uso1p/p115保守。UsoA与粗糙脉孢霉、酿酒酵母的Uso1和p115的同源性分别为46%、29%、29%和26%。庞贝和牛肉。我们重点研究了通过抑制usoA基因在真菌细胞内的蛋白运输。

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.