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编码诱导型酶分解代谢脱氢奎尼酸酶的粗糙脉孢菌结构基因在大肠杆菌中的组成型表达。

Constitutive expression in Escherichia coli of the Neurospora crassa structural gene encoding the inducible enzyme catabolic dehydroquinase.

作者信息

Hautala J A, Schweizer M, Giles N H, Kushner S R

出版信息

Mol Gen Genet. 1979 Apr 17;172(1):93-8. doi: 10.1007/BF00276219.

Abstract

In Neurospora crassa the qa-2 gene, which encodes catabolic dehydroquinase, is under positive control exerted by the inducer quinic acid and an activator protein encoded in the closely linked qa-1 gene. In order to determine if this regulatory mechanism is maintained when the qa-2 gene is cloned on a recombinant plasmid and expressed in Escherichia coli, molecular cloning experiments have been performed using DNA isolated from a qa-1+ (inducible), a qa-1C (constitutive) and two qa-1 (non-inducible) strains of N. crassa. The results demonstrate that the level of expression of the qa-2 gene in E. coli is completely independent of the mutational state of the qa-1 gene. Moreover, the level of expression of the cloned qa-2 gene was unaffected by either an intracellularly produced inducer of catabolic dehydroquinase or by the general procaryotic positive effector, the CAP factor. The weight of evidence thus supports the conclusion that transcription of the N. crassa qa-2 gene in E. coli does not require the qa-1 activator protein and thus is not controlled by the same mecahnism which functions in N. crassa.

摘要

在粗糙脉孢菌中,编码分解代谢脱氢奎尼酸酶的qa - 2基因受诱导物奎尼酸和紧密连锁的qa - 1基因编码的激活蛋白的正向调控。为了确定当qa - 2基因克隆到重组质粒上并在大肠杆菌中表达时这种调控机制是否仍然存在,已经使用从粗糙脉孢菌的一个qa - 1 +(可诱导型)、一个qa - 1C(组成型)和两个qa - 1(不可诱导型)菌株中分离的DNA进行了分子克隆实验。结果表明,qa - 2基因在大肠杆菌中的表达水平完全独立于qa - 1基因的突变状态。此外,克隆的qa - 2基因的表达水平不受分解代谢脱氢奎尼酸酶的细胞内产生的诱导物或原核生物通用正向效应物CAP因子的影响。因此,大量证据支持这样的结论:粗糙脉孢菌qa - 2基因在大肠杆菌中的转录不需要qa - 1激活蛋白,因此不受在粗糙脉孢菌中起作用的相同机制的控制。

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