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包含米曲霉淀粉酶A启动子和大肠杆菌β-葡萄糖醛酸酶基因的融合基因的构建及其在米曲霉中的表达分析。

Construction of a fusion gene comprising the Taka-amylase A promoter and the Escherichia coli beta-glucuronidase gene and analysis of its expression in Aspergillus oryzae.

作者信息

Tada S, Gomi K, Kitamoto K, Takahashi K, Tamura G, Hara S

机构信息

Research Institute of Brewing Resources Co. Ltd., Tokyo, Japan.

出版信息

Mol Gen Genet. 1991 Oct;229(2):301-6. doi: 10.1007/BF00272170.

Abstract

Northern blot analysis of glucose-grown and starch-grown mycelia of Aspergillus oryzae RIB40 was conducted using the cloned Taka-amylase A (TAA) gene as a probe. The amount of mRNA homologous to the TAA gene was increased when this fungus was grown with starch as a sole carbon source. In order to analyze the induction mechanism, we inserted the Escherichia coli uidA gene encoding beta-glucuronidase (GUS) down-stream of the TAA promoter and introduced the resultant fusion gene into the A. oryzae genome. Production of a functional GUS protein was induced by starch, but not by glucose. When the effects of various sugars on expression of the fusion gene were examined, the results suggested that the expression of the fusion gene was under control of the TAA gene promoter.

摘要

使用克隆的米曲霉淀粉酶A(TAA)基因作为探针,对米曲霉RIB40以葡萄糖为碳源生长的菌丝体和以淀粉为碳源生长的菌丝体进行 Northern 印迹分析。当这种真菌以淀粉作为唯一碳源生长时,与TAA基因同源的mRNA量增加。为了分析诱导机制,我们将编码β-葡萄糖醛酸酶(GUS)的大肠杆菌uidA基因插入TAA启动子的下游,并将所得融合基因导入米曲霉基因组。功能性GUS蛋白的产生受淀粉诱导,但不受葡萄糖诱导。当检测各种糖类对融合基因表达的影响时,结果表明融合基因的表达受TAA基因启动子的控制。

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