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大肠杆菌K12中参与磷酸盐限制诱导蛋白合成调控的phoM基因的克隆。

Cloning of phoM, a gene involved in regulation of the synthesis of phosphate limitation inducible proteins in Escherichia coli K12.

作者信息

Tommassen J, Heimstra P, Overduin P, Lugtenberg B

出版信息

Mol Gen Genet. 1984;195(1-2):190-4. doi: 10.1007/BF00332745.

Abstract

Like the synthesis of alkaline phosphatase, the synthesis of outer membrane PhoE protein is shown to be dependent on the phoM gene product in phoR mutants of E. coli K12. This phoM gene has been cloned into the multi-copy vector pACYC184 using selection for alkaline phosphatase constitutive synthesis in a phoR background. The gene was localized on the hybrid plasmids by analysis of deletion plasmids constructed in vitro and of mutant plasmids generated by gamma delta insertions. Interestingly, two of the selected hybrid plasmids contained the entire phoA-phoB-phoR region of the chromosome, as a multiple copy state of these genes results in the constitutive synthesis of alkaline phosphatase. The presence of multiple copies of the phoM gene hardly influences the level of expression of alkaline phosphatase and PhoE protein in a pho+ strain, but significantly increases the levels of these proteins in a phoR mutant strain.

摘要

与碱性磷酸酶的合成一样,在大肠杆菌K12的phoR突变体中,外膜PhoE蛋白的合成显示依赖于phoM基因产物。利用在phoR背景下对碱性磷酸酶组成型合成的选择,已将该phoM基因克隆到多拷贝载体pACYC184中。通过分析体外构建的缺失质粒和由γδ插入产生的突变体质粒,将该基因定位在杂交质粒上。有趣的是,两个选定的杂交质粒包含染色体的整个phoA-phoB-phoR区域,因为这些基因的多拷贝状态导致碱性磷酸酶的组成型合成。phoM基因多拷贝的存在对pho+菌株中碱性磷酸酶和PhoE蛋白的表达水平几乎没有影响,但显著增加了phoR突变菌株中这些蛋白的水平。

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