Abstract
A reporter construct was created on the basis of the transcription attenuator region of the Escherichia coli tryptophan operon. Dual-fluorescent-protein genes for red fluorescent protein and cerulean fluorescent protein were used as a sensor and internal control of gene expression. The sequence of the attenuator was modified to avoid tryptophan sensitivity while preserving sensitivity to ribosome stalling. Antimicrobial compounds which cause translation arrest at the stage of elongation induce the reporter both in liquid culture and on an agar plate. This reporter could be used for high-throughput screening of translation inhibitors.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Agar
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Amino Acid Sequence
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Anti-Bacterial Agents / biosynthesis
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Anti-Bacterial Agents / pharmacology
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Culture Media
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Escherichia coli / metabolism
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Fermentation
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Gene Expression Regulation, Bacterial / drug effects
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Genes, Reporter / genetics
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High-Throughput Screening Assays / methods*
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Luminescent Proteins / genetics*
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Micromonospora / metabolism
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Molecular Sequence Data
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Operon
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Peptide Chain Elongation, Translational / drug effects*
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Protein Synthesis Inhibitors / pharmacology
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Red Fluorescent Protein
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Ribosomes / drug effects
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Siphoviridae / genetics
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Transcription, Genetic / drug effects*
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Tryptophan / genetics
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Tryptophan / pharmacology
Substances
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Anti-Bacterial Agents
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Culture Media
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Luminescent Proteins
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Protein Synthesis Inhibitors
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Tryptophan
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Agar