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过量表达的大肠杆菌katF/rpoS基因产物的体外功能特性分析

In vitro functional characterization of overproduced Escherichia coli katF/rpoS gene product.

作者信息

Nguyen L H, Jensen D B, Thompson N E, Gentry D R, Burgess R R

机构信息

McArdle Laboratory for Cancer Research, University of Wisconsin-Madison 53706.

出版信息

Biochemistry. 1993 Oct 19;32(41):11112-7. doi: 10.1021/bi00092a021.

Abstract

The katF/rpoS gene product (sigma s), a central regulator of stationary-phase gene expression in Escherichia coli, has been purified from an overproducing strain. sigma s was used as an immunogen for the production of monoclonal antibodies. Previous sequence analysis of sigma s strongly indicated homology to the sigma factor family. We show biochemically in this paper that sigma s is a sigma factor. This protein can bind to core RNA polymerase (E), and this binding can be competed effectively by the major E. coli transcription initiation factor, sigma 70. Immunopurified sigma s holoenzyme (E sigma s) transcribes the promoters of the bolAp1 gene and the xthA gene. Interestingly, both promoters can also be transcribed by sigma 70 holoenzyme (E sigma 70).

摘要

katF/rpoS基因产物(σs)是大肠杆菌中稳定期基因表达的核心调节因子,已从高产菌株中纯化出来。σs被用作免疫原以制备单克隆抗体。先前对σs的序列分析强烈表明其与σ因子家族具有同源性。我们在本文中通过生化方法表明σs是一种σ因子。这种蛋白质可以与核心RNA聚合酶(E)结合,并且大肠杆菌主要转录起始因子σ70可以有效地竞争这种结合。免疫纯化的σs全酶(Eσs)可转录bolAp1基因和xthA基因的启动子。有趣的是,这两个启动子也可以被σ70全酶(Eσ70)转录。

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