Hisada Hiromoto, Sano Motoaki, Hata Yoji, Abe Yasuhisa, Machida Masayuki
Research Institute, Gekkeikan Sake Co., Ltd., Kyoto, Japan.
Biosci Biotechnol Biochem. 2008 Jan;72(1):48-53. doi: 10.1271/bbb.70321. Epub 2008 Jan 7.
The catalase-encoding gene (catB) is expressed strongly in Aspergillus oryzae. To identify the transcription regulatory elements involved in strong expression, we did promoter deletion analysis using beta-glucuronidase (GUS) as a reporter and an electrophoretic gel mobility shift assay (EMSA) systematically. The deletion 200-bp sequence from -1,000 to -800 in the 1,400-bp catB promoter caused a drastic decrease in GUS activity. In addition, EMSA implicated a 45-bp element from -1,000 to -956 containing cis-elements. According to detailed promoter deletion analysis, a region from -1,000 to -975, which contains putative heat shock element (HSE) and the CCAAT-box, was involved in strong expression.
过氧化氢酶编码基因（catB）在米曲霉中强烈表达。为了鉴定参与强表达的转录调控元件，我们系统地使用β-葡萄糖醛酸酶（GUS）作为报告基因进行启动子缺失分析，并进行了电泳凝胶迁移率变动分析（EMSA）。在1400 bp的catB启动子中，从-1000到-800缺失200 bp序列导致GUS活性急剧下降。此外，EMSA表明从-1000到-956的一个45 bp元件含有顺式元件。根据详细的启动子缺失分析，包含假定热休克元件（HSE）和CCAAT框的-1000到-975区域参与了强表达。
