Tsuchiya K, Tada S, Gomi K, Kitamoto K, Kumagai C, Tamura G
Research Institute of Brewing Resources Co., Ltd., Tokyo, Japan.
Biosci Biotechnol Biochem. 1992 Nov;56(11):1849-53. doi: 10.1271/bbb.56.1849.
The Taka-amylase A gene (amyB) of Aspergillus oryzae is induced by starch or maltose. The molecular mechanism of the induction was investigated using a fusion of the amyB promoter and the Escherichia coli uidA gene encoding beta-glucuronidase (GUS). To identify the region responsible for high-level expression and regulation within the amyB promoter, a series of deletion promoters was constructed and introduced into the A. oryzae met locus by homologous recombination. Deletion of the region between -377 to -290 (the number indicates the distance in base pairs from the translation initiation point (+1) to the deletion end point) significantly reduced of the GUS activity, but slight reduction of the GUS activity was observed in deletions up to -377. Northern blot analysis showed that reduction of the GUS activity depended upon the expression level of the GUS gene. The region between -377 to -290 is suggested to include the sequence required directly for high-level expression and regulation of the amyB gene.
米曲霉的Taka淀粉酶A基因（amyB）由淀粉或麦芽糖诱导。利用amyB启动子与编码β-葡萄糖醛酸酶（GUS）的大肠杆菌uidA基因的融合体，研究了诱导的分子机制。为了鉴定amyB启动子内负责高水平表达和调控的区域，构建了一系列缺失启动子，并通过同源重组将其导入米曲霉的met位点。缺失-377至-290之间的区域（数字表示从翻译起始点（+1）到缺失终点的碱基对距离）显著降低了GUS活性，但在缺失至-377的情况下，GUS活性略有降低。Northern印迹分析表明，GUS活性的降低取决于GUS基因的表达水平。-377至-290之间的区域被认为包含amyB基因高水平表达和调控直接所需的序列。
