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一种与其他细菌的阻遏系统同源的阻遏系统在天蓝色链霉菌精氨酸生物合成基因控制中的作用。

Implication of a repression system, homologous to those of other bacteria, in the control of arginine biosynthesis genes in Streptomyces coelicolor.

作者信息

Soutar A, Baumberg S

机构信息

Department of Genetics, University of Leeds, UK.

出版信息

Mol Gen Genet. 1996 May 23;251(2):245-51. doi: 10.1007/BF02172924.

Abstract

As with most amino acid biosynthetic pathways in streptomycetes, enzymes of arginine biosynthesis in Streptomyces coelicolor show only slight derepression in minimal medium without, as opposed to with, exogenous arginine. However, when an arginine auxotroph was cultured in limiting arginine, ornithine carbamoyltransferase (OCT) activities rose by as much as 100-fold. The response was not due to a general starvation effect. To elucidate the repression-derepression mechanism, a DNA fragment containing the upstream region of the previously isolated S. coelicolor argCJB cluster was cloned into a multicopy vector and transformed into wild-type S. coelicolor; a slight transient derepression of OCT was observed in minimal medium without, though not with, added arginine, consistent with titration by the insert of a negatively acting macromolecule such as a repressor. A subfragment carrying the 5' end of argC and the region immediately upstream showed specific binding, in mobility shift assays, to purified AhrC, the repressor/activator of genes of arginine metabolism in Bacillus subtilis. It is therefore likely that in S. coelicolor, expression of arginine biosynthesis genes is controlled by a protein homologous to the well-characterised B. subtilis and Escherichia coli repressors.

摘要

与链霉菌中大多数氨基酸生物合成途径一样,在不含(与含相对)外源精氨酸的基本培养基中,天蓝色链霉菌精氨酸生物合成的酶仅表现出轻微的去阻遏。然而,当精氨酸营养缺陷型在限量精氨酸中培养时,鸟氨酸氨甲酰基转移酶(OCT)活性可提高多达100倍。这种反应并非由于普遍的饥饿效应。为阐明阻遏-去阻遏机制,将包含先前分离的天蓝色链霉菌argCJB簇上游区域的DNA片段克隆到多拷贝载体中,并转化到野生型天蓝色链霉菌中;在不添加(而非添加)精氨酸的基本培养基中观察到OCT有轻微的短暂去阻遏,这与插入负性作用大分子(如阻遏物)进行滴定一致。在迁移率变动分析中,携带argC 5'端和紧邻上游区域的一个亚片段显示与纯化的AhrC特异性结合,AhrC是枯草芽孢杆菌精氨酸代谢基因的阻遏物/激活物。因此,在天蓝色链霉菌中,精氨酸生物合成基因的表达可能由与特征明确的枯草芽孢杆菌和大肠杆菌阻遏物同源的一种蛋白质控制。

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