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噬菌体P1的c1阻遏物失活蛋白coi。coi的克隆与表达及其对c1阻遏物功能的干扰。

The c1 repressor inactivator protein coi of bacteriophage P1. Cloning and expression of coi and its interference with c1 repressor function.

作者信息

Heinzel T, Velleman M, Schuster H

机构信息

Max-Planck Institut für Molekulare Genetik, Berlin, Federal Republic of Germany.

出版信息

J Biol Chem. 1990 Oct 15;265(29):17928-34.

PMID:2211669
Abstract

The immC region of bacteriophage P1 contains the c1 repressor gene and its upstream region with four c1-controlled operators and four open reading frames. A c1 inactivator gene, coi, was defined by mutations in immC that suppress the virulence of the P1virC mutation. The exact location of the coi gene was not known (Scott, J.R. (1980) Curr. Top. Microbiol. Immunol. 90, 49-65). When a variety of P1 immC fragments were inserted into an expression vector, a gene product was inducible for the open reading frame 4 only. We identify this product as the c1 inactivator protein, coi by the following criteria: (a) expression of coi from a recombinant plasmid induces the P1 prophage and inhibits lysogenization of sensitive bacteria by P1; (b) all c1-controlled operator-promoter elements tested in vivo are derepressed by coi; (c) a partially purified coi protein (apparent molecular weight = 4800) interacts with c1 repressor and inhibits its binding to the operator in vitro. Based on these results we refine a model for the regulation of those genes and elements within immC which participate in the decision of P1 to enter the lytic or lysogenic pathway.

摘要

噬菌体P1的immC区域包含c1阻遏基因及其上游区域,该上游区域有四个受c1控制的操纵子和四个开放阅读框。一个c1失活基因coi,是通过immC中的突变来定义的,这些突变可抑制P1virC突变的毒力。coi基因的确切位置尚不清楚(斯科特,J.R.(1980年)《微生物学与免疫学当前专题》90,49 - 65)。当将各种P1 immC片段插入到一个表达载体中时,一种基因产物仅对开放阅读框4具有诱导性。我们通过以下标准将该产物鉴定为c1失活蛋白coi:(a)来自重组质粒的coi表达可诱导P1原噬菌体,并抑制P1对敏感细菌的溶源化;(b)在体内测试的所有受c1控制的操纵子 - 启动子元件都被coi去阻遏;(c)一种部分纯化的coi蛋白(表观分子量 = 4800)在体外与c1阻遏物相互作用并抑制其与操纵子的结合。基于这些结果,我们完善了一个模型,用于调控immC内那些参与P1进入裂解或溶源途径决策的基因和元件。

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