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SUC2基因的上游区域赋予酿酒酵母中异源基因以调控表达。

Upstream region of the SUC2 gene confers regulated expression to a heterologous gene in Saccharomyces cerevisiae.

作者信息

Sarokin L, Carlson M

出版信息

Mol Cell Biol. 1985 Oct;5(10):2521-6. doi: 10.1128/mcb.5.10.2521-2526.1985.

Abstract

The SUC2 gene produces two differently regulated mRNAs that encode two forms of invertase. The 1.9-kilobase mRNA encoding secreted invertase is regulated by glucose (carbon catabolite) repression, and the 1.8-kilobase mRNA encoding intracellular invertase is synthesized constitutively. Previous work has shown that the 5' noncoding region between -650 and -418 is required for derepression of secreted invertase in response to glucose deprivation. We show here that this upstream region can confer glucose-repressible expression to a heterologous gene, a LEU2-lacZ gene fusion, that is not normally regulated by glucose repression. This expression was found to respond appropriately to mutations in trans-acting genes that affect regulation of SUC2 expression. Mutations in the SNF1 through SNF6 loci reduced derepression of beta-galactosidase, and a mutation at the SSN6 locus caused constitutive expression. These findings indicate that the SUC2 upstream region mediates the regulatory effects of these genes and suggest that regulation occurs at the level of transcription. In addition, the upstream region was partially active in the inverted orientation.

摘要

SUC2基因产生两种受不同调控的mRNA,它们编码两种形式的转化酶。编码分泌型转化酶的1.9千碱基mRNA受葡萄糖(碳代谢物)阻遏调控,而编码细胞内转化酶的1.8千碱基mRNA则组成型合成。先前的研究表明,-650至-418之间的5'非编码区是分泌型转化酶在葡萄糖缺乏时去阻遏所必需的。我们在此表明,该上游区域可赋予一个异源基因(LEU2-lacZ基因融合体)葡萄糖可阻遏的表达,该基因通常不受葡萄糖阻遏调控。发现这种表达对影响SUC2表达调控的反式作用基因突变有适当反应。SNF1至SNF6位点的突变降低了β-半乳糖苷酶的去阻遏作用,而SSN6位点的突变导致组成型表达。这些发现表明,SUC2上游区域介导了这些基因的调控作用,并提示调控发生在转录水平。此外,上游区域在反向排列时部分具有活性。

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