Lee B R, Kitamoto K, Yamada O, Kumagai C
National Research Institute of Brewing, Tokyo, Japan.
Appl Microbiol Biotechnol. 1995 Dec;44(3-4):425-31. doi: 10.1007/BF00169939.
The nuclease S1 gene (nucS) from Aspergillus oryzae was isolated using a polymerase-chain-reaction-amplified DNA fragment as a probe, and a 2.6-kb SalI-EcoRI fragment containing the nucS gene was sequenced. It was deduced that the nucS gene had two short introns, 49 and 50 nucleotides in length. The nucS gene had an open-reading frame of 963 base pairs and coded for a protein of 287 amino acid residues, comprising the signal peptide of 20 amino acids and a mature protein of 267 amino acids. The deduced amino acid sequence agreed well with the published amino acid sequence except for one substitution. Southern hybridization analysis showed that the nucS gene existed as a single copy in the A. oryzae chromosome. When the structural gene of nucS was fused with the promoter of the glaA gene and introduced into A. oryzae, the yield of secreted nuclease S1 increased about 100-fold compared with the recipient strain.
以聚合酶链反应扩增的DNA片段为探针，从米曲霉中分离出核酸酶S1基因（nucS），并对包含nucS基因的2.6kb SalI-EcoRI片段进行了测序。推断nucS基因有两个短内含子，长度分别为49和50个核苷酸。nucS基因有一个963个碱基对的开放阅读框，编码一个由287个氨基酸残基组成的蛋白质，包括20个氨基酸的信号肽和267个氨基酸的成熟蛋白。推导的氨基酸序列与已发表的氨基酸序列除一处替换外吻合良好。Southern杂交分析表明，nucS基因在米曲霉染色体中以单拷贝形式存在。当nucS的结构基因与glaA基因的启动子融合并导入米曲霉时，分泌型核酸酶S1的产量与受体菌株相比增加了约100倍。
