Campbell Jennifer
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts.
Curr Protoc Chem Biol. 2010 Dec 1;2(4):195-208. doi: 10.1002/9780470559277.ch100115.
The increasing incidence of antibiotic-resistant bacterial infections both in hospitals and in the community intensifies the need for new antibacterial strategies and targets. Although high-throughput screening against live bacteria allows rapid discovery of compounds with growth-inhibitory activities, these efforts have failed to fill the pipeline with the anticipated antibacterial compounds because target identification is often onerous. Recently, a strategy was reported that employs a bacterial growth inhibition assay readout using optical density measurements on paired strains-both a wild-type strain and a pathway-null mutant-to find inhibitors of wild-type bacterial growth that specifically target conditionally essential enzymes in the pathway of interest. Protocols are provided here for determining the robustness of an assay, screening in a high-throughput format, and setting up dose-response curves in paired Staphylococcus aureus strains. However, the protocols can be used to screen for growth-inhibitory compounds in any bacterial strain of interest. Curr. Protoc. Chem. Biol. 2:195-208 © 2010 by John Wiley & Sons, Inc.
医院和社区中抗生素耐药性细菌感染的发生率不断上升，这加剧了对新抗菌策略和靶点的需求。尽管针对活细菌的高通量筛选能够快速发现具有生长抑制活性的化合物，但由于靶点鉴定通常很繁琐，这些努力未能使预期的抗菌化合物充实研发渠道。最近，有报道称一种策略利用了细菌生长抑制试验读数，该读数通过对配对菌株（野生型菌株和途径缺失突变体）进行光密度测量来寻找野生型细菌生长的抑制剂，这些抑制剂专门针对感兴趣途径中的条件必需酶。本文提供了用于确定试验稳健性、高通量筛选以及在配对金黄色葡萄球菌菌株中建立剂量反应曲线的方案。然而，这些方案可用于筛选任何感兴趣细菌菌株中的生长抑制化合物。《当代化学与生物学实验指南》2:195 - 208 © 2010 约翰威立父子公司
