Deletion analysis of promoter elements of the Aspergillus oryzae agdA encoding alpha-glucosidase
Deletion analysis of promoter elements of the Aspergillus oryzae agdA encoding alpha-glucosidase
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DOI：
10.1007/s002940050153
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发表时间：
1996-11-01
期刊：
影响因子：
2.5
通讯作者：
Tamura, G
中科院分区：
文献类型：
--
作者：
Minetoki, T;Nunokawa, Y;Tamura, G
测定了编码α-葡糖苷酶的曲霉菌agdA基因的启动子区的1.5-kb片段的核苷酸序列。与其他曲霉淀粉酶基因启动子区的比较表明，有三个高度保守的序列，命名为区域I，II和III，分别位于相对于起始密码子的-670 nt，-596 nt和-544 nt。使用niaD同源转化系统，通过缺失与大肠杆菌uidA基因融合的启动子分析，研究agdA启动子中这些共有序列的功能，缺失区域III的上游半部分（IIIa;-544至-529）导致GUS活性降低超过90%并且消除麦芽糖诱导，这表明区域IIIa是高水平表达和麦芽糖诱导的功能性必需元件，区域I和区域III的下游一半的缺失（IIIb;-521至-511）导致GUS活性显著降低，但不影响麦芽糖诱导，这表明，这两个元件最有可能包含与区域IIIa合作参与有效表达的序列。此外，区域IIIb和推定的TATA bos之间的340 bp区域的缺失导致活性增加2倍。
The nucleotide sequence of a 1.5-kb fragment of the promoter region of the Aspergillus oryzae agdA gene encoding alpha-glucosidase was determined. A comparison with the promoter regions of other Aspergillus amylase genes indicated that there are three highly conserved sequences, designated Regions I, II and III, located at - 670 nt, - 596 nt and - 544 nt relative to the start codon, respectively. The function of these consensus sequences in the agdA promoter was investigated by deletion analysis of a promoter fusion with the Escherichia coli uidA gene, using the niaD homologous-transformation system, Deletion of the upstream half of Region III (IIIa; - 544 to - 529) resulted in a more than 90% reduction in GUS activity and abolished maltose induction, suggesting that Region IIIa is a functionally essential element for high-level expression and maltose induction, Deletion of Region I and the downstream half of Region III (IIIb; - 521 to - 511) resulted in a significant reduction in GUS activity, but did not affect maltose induction, This suggested that these two elements most likely contain sequences involved in efficient expression in cooperation with Region IIIa. In addition, deletion of a 340-bp region between Region IIIb and the putative TATA bos resulted in a 2-fold increase in activity.