Whole cell strategies based on lux genes for high throughput applications toward new antimicrobials.
The discovery/development of novel drug candidates has witnessed dramatic changes over the last two decades. Old methods to identify lead compounds are not suitable to screen wide libraries generated by combinatorial chemistry techniques. High throughput screening (HTS) has become irreplaceable and hundreds of different approaches have been described. Assays based on purified components are flanked by whole cell-based assays, in which reporter genes are used to monitor, directly or indirectly, the influence of a chemical over the metabolism of living cells. The most convenient and widely used reporters for real-time measurements are luciferases, light emitting enzymes from evolutionarily distant organisms. Autofluorescent proteins have been also extensively employed, but proved to be more suitable for end-point measurements, in situ applications - such as the localization of fusion proteins in specific subcellular compartments - or environmental studies on microbial populations. The trend toward miniaturization and the technical advances in detection and liquid handling systems will allow to reach an ultra high throughput screening (uHTS), with 100,000 of compounds routinely screened each day. Here we show how similar approaches may be applied also to the search for new and potent antimicrobial agents.
Galluzzi
Lorenzo
L
Institute of Environmental Engineering and Biotechnology, Tampere University of Technology, P.O. Box 541, FIN-33101 Tampere, Finland.
Karp
Matti
M
D000890:Anti-Infective Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001681:Biological Assay / Q000191:economics / Q000379:methods*; D015374:Biosensing Techniques / Q000191:economics / Q000379:methods*; D020650:Combinatorial Chemistry Techniques / Q000191:economics / Q000379:methods; D003584:Cytological Techniques; D004353:Drug Evaluation, Preclinical / Q000379:methods; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D049449:Luminescence; D011387:Prokaryotic Cells / Q000166:cytology / Q000378:metabolism
D000890:Anti-Infective Agents; D001426:Bacterial Proteins; D008156:Luciferases