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米曲霉过氧化氢酶编码基因(catB)启动子的缺失分析

Deletion analysis of the catalase-encoding gene (catB) promoter from Aspergillus oryzae.

作者信息

Hisada Hiromoto, Sano Motoaki, Hata Yoji, Abe Yasuhisa, Machida Masayuki

机构信息

Research Institute, Gekkeikan Sake Co., Ltd., Kyoto, Japan.

出版信息

Biosci Biotechnol Biochem. 2008 Jan;72(1):48-53. doi: 10.1271/bbb.70321. Epub 2008 Jan 7.

Abstract

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.

摘要

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。

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