A Pathway-specific Cell Based Screening System to Detect Bacterial Cell Wall Inhibitors.

DOI PubMed 被引用文献4件 参考文献31件

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A pathway-specific cell-based screen is described to detect compounds that inhibit the biosynthesis of the cell wall of bacteria. The basis for detection is the discovery that the β-lactamase gene from Citrobacter freundii, cloned into Escherichia coli, is induced when cells are exposed to known cell wall inhibitors, and not just β-lactam-based antibiotics. In a wild type host, cell wall inhibitors such as moenomycin, vancomycin, and ramoplanin, which are excluded by the outer membrane, only induce at high concentrations. However, these compounds, as well as fosfomycin, cycloserine, and cefoxitin, induce at concentrations at or below the MIC of a host carrying the envA-mutation, which causes a defect in the outer membrane. As additional proof that induction of β-lactamase is the direct result of cell wall inhibition, a host strain carrying a temperature-sensitive mutation in the murG gene, whose product converts the cell wall intermediate Lipid I, to Lipid II, also induced β-lactamase at the restrictive temperature. A protocol is described for screening samples in high-throughput mode.

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