title: Attenuation-based dual-fluorescent-protein reporter for screening translation inhibitors.
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.
authors.affiliation: Department of Chemistry and A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia.
authors.forename: Ilya A
authors.initials: IA
authors.lastname: Osterman
authors.forename: Irina V
authors.initials: IV
authors.lastname: Prokhorova
authors.forename: Vasily O
authors.initials: VO
authors.lastname: Sysoev
authors.forename: Yulia V
authors.initials: YV
authors.lastname: Boykova
authors.forename: Olga V
authors.initials: OV
authors.lastname: Efremenkova
authors.forename: Maxim S
authors.initials: MS
authors.lastname: Svetlov
authors.forename: Vyacheslav A
authors.initials: VA
authors.lastname: Kolb
authors.forename: Alexey A
authors.initials: AA
authors.lastname: Bogdanov
authors.forename: Petr V
authors.initials: PV
authors.lastname: Sergiev
authors.forename: Olga A
authors.initials: OA
authors.lastname: Dontsova
mesh_terms: D000362:Agar; D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000096:biosynthesis / Q000494:pharmacology; D003470:Culture Media; D004926:Escherichia coli / Q000378:metabolism; D005285:Fermentation; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D017930:Genes, Reporter / Q000235:genetics; D057166:High-Throughput Screening Assays / Q000379:methods*; D008164:Luminescent Proteins / Q000235:genetics*; D008847:Micromonospora / Q000378:metabolism; D008969:Molecular Sequence Data; D009876:Operon; D010441:Peptide Chain Elongation, Translational / Q000187:drug effects*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012270:Ribosomes / Q000187:drug effects; D017906:Siphoviridae / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects*; D014364:Tryptophan / Q000235:genetics / Q000494:pharmacology; D000097573:Red Fluorescent Protein
chemical_list: D000362:Agar; D000900:Anti-Bacterial Agents; D003470:Culture Media; D008164:Luminescent Proteins; D011500:Protein Synthesis Inhibitors; D014364:Tryptophan; D000097573:Red Fluorescent Protein