Design of a Novel Mammalian Screening System for the Detection of Bioavailable, Non-cytotoxic Streptogramin Antibiotics.
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- AUBEL DOMINIQUE
- Institut Universitaire de Technologic, IUTA, Département Génie Biologique
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- MORRIS ROWAN
- Biozentrum, University of Basel, Department of Microbiology
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- LENNON BARBARA
- Institute of Biotechnology, Swiss Federal Institute of Technology
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- RIMANN MARKUS
- Institute of Biotechnology, Swiss Federal Institute of Technology
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- KAUFMANN HITTO
- Institute of Biotechnology, Swiss Federal Institute of Technology
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- FOLCHER MARC
- Biozentrum, University of Basel, Department of Microbiology
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- BAILEY JAMES E.
- Institute of Biotechnology, Swiss Federal Institute of Technology
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- THOMPSON CHARLES J.
- Biozentrum, University of Basel, Department of Microbiology
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- FUSSENEGGER MARTIN
- Institute of Biotechnology, Swiss Federal Institute of Technology
抄録
Screening and development of new antibiotic activities to counteract the increasing prevalence of multidrug-resistant (MDR) human pathogenic bacteria has once again become a priority in human chemotherapy. Here we describe a novel mammalian cell culture-based screening platform for the detection of Streptogramin antibiotics. Quinupristin-dalfopristin (Synercid®), a synthetically modified Streptogramin, is presently the sole effective agent in the treatment of some MDR nosocomial infections. A Streptomyces coelicolor transcriptional regulator (Pip) has been adapted to modulate reporter gene expression (SEAP, secreted alkaline phosphatase) in Chinese hamster ovary cells (CHO) in response to Streptogramin antibiotics. This CHO cell-based technology was more sensitive in detecting the production of the model Streptogramin pristinamycin, from Streptomyces pristinaespiralis, than antibiogram tests using a variety of human pathogenic bacteria as indicator strains. The reporter system was able to detect pristinamycin compound produced by a single S. pristinaespiralis colony. The assay was rapid (17 hours) and could be carried out in a high-throughput 96-well plate assay format or a 24-well transwell set-up. This novel mammalian cell-based antibiotic screening concept enables detection of bioavailable and non-cytotoxic representatives of a particular class of antibiotics in a single assay and represents a promising alternative to traditional antibiogram-based screening programs.
収録刊行物
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- The Journal of Antibiotics
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The Journal of Antibiotics 54 (1), 44-55, 2001
公益財団法人 日本感染症医薬品協会