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利用phoB-氯霉素乙酰转移酶融合基因分析phoB表达的调控

Analysis of regulation of phoB expression using a phoB-cat fusion.

作者信息

Guan C D, Wanner B, Inouye H

出版信息

J Bacteriol. 1983 Nov;156(2):710-7. doi: 10.1128/jb.156.2.710-717.1983.

Abstract

The phoB gene, which encodes a positive control factor for a number of phosphate-regulated genes in Escherichia coli, was cloned into multicopy plasmid pBR322. A phoB-cat fusion that expressed chloramphenicol transacetylase from the phoB promoter was constructed. Studies of the expression of the phoB-cat fusion showed that the pattern of regulation of the phoB gene was similar to that of the phoA gene, the structural gene for alkaline phosphatase. The phoB gene was derepressed under conditions of phosphate starvation, was constitutively expressed in a phoR background, and required the phoM gene product for expression in a phoR strain. Finally, a functional phoB product was required for its own synthesis. Our results indicate either that phoA gene expression responds directly to the concentration of the phoB gene product in cells or that the phoA and phoB controlling elements are quite similar.

摘要

编码大肠杆菌中多个磷酸盐调节基因的正调控因子的phoB基因,被克隆到多拷贝质粒pBR322中。构建了一个从phoB启动子表达氯霉素转乙酰酶的phoB-cat融合体。对phoB-cat融合体表达的研究表明,phoB基因的调控模式与碱性磷酸酶结构基因phoA基因的调控模式相似。phoB基因在磷酸盐饥饿条件下被去阻遏,在phoR背景中组成型表达,并且在phoR菌株中表达需要phoM基因产物。最后,其自身的合成需要功能性的phoB产物。我们的结果表明,要么phoA基因表达直接响应细胞中phoB基因产物的浓度,要么phoA和phoB的控制元件非常相似。

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