Project description:6 benign TURP samples taken directly from patients were subjected to collagen digestion overnight. Epithelial cells isolated were adhered to collagen for 15 minutes. Adherent cells were passed through a magnetic bead column and cells positive for CD133 antibody were double selected. RNA was extracted from CD133+ and CD133- cells from each patient and amplified using the RiboAmp HS RNA amplification system. CD133+ (Cy5) and CD133- (Cy5) cDNA was hybridised to 30K cDNA array (CRUK facility) using the Invitrogen post labelling system. In each case that RNA was co-hybridised to a human pooled universal reference cDNA (Cy3). Keywords: repeat sample
Project description:Antimicrobial resistance is a leading mortality factor worldwide. Here we report the discovery of clovibactin, a new antibiotic, isolated from uncultured soil bacteria. Clovibactin efficiently kills drug-resistant Gram-positivebacterial pathogens without detectable resistance. Using biochemical assays,solid-state NMR, and atomic force microscopy, we dissect its mode of action. Clovibactin blocks cell wall synthesis by targeting pyrophosphate of multiple essential peptidoglycan precursors (C55PP, Lipid II, LipidWTA). Clovibactin uses anunusual hydrophobic interface to tightly wrap aroundpyrophosphate, butbypasses the variable structural elements of precursors, accounting for the lack of resistance. Selective and efficient target binding is achieved by the sequestration of precursors into supramolecular fibrils that only form on bacterial membranes that contain lipid-anchored pyrophosphate groups.Uncultured bacteria offer a rich reservoir of antibiotics with new mechanisms of action that could replenish the antimicrobial discovery pipeline.
