Before sequencing, the PCR products were purified using QIAquick

PCR purification kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions. Isolation and analysis of LPS LPS was isolated and analyzed by a two-buffer tricine-based SDS-PAGE system. The isolation of the LPS was performed as described previously [16]. The SDS-PAGE consists of a 4% stacking gel and a 16.5% separating gel. Before analysis by SDS-PAGE, an aliquot of the LPS sample was combined with an equal volume of 2 × sample buffer (0.2% bromophenol blue, 10% β-mercaptoethanol, 40% glycerol, 3.3% SDS and 100 mM Tris HCL, pH6.8) and heated to 95°C for 5 min. Before silver staining with 0.1% silver nitrate, the learn more gels were incubated in acetic acid for 30 min. After 5 min washing in dH2O, the gels were developed in 2.5% sodium carbonate, 0.1% formaldehyde, 0.001% sodiumthiosulfate for 2-5 min. To stop the reaction, the gels were transferred into a 2% glycine, 0.5% EDTA solution. Identification of Savolitinib in vivo promoter regions, terminator

structures and other motifs The genome of phage JG004 was scanned for the presence of putative sigma 70-dependent promoter regions using the web service SAK [22]. Putative promoter regions with a score above 1 were scanned for the presence of https://www.selleckchem.com/products/AZD8931.html conserved -10 and -35 regions using the Virtual Footprint software [53] and for their genomic location, orientation and vicinity to the next gene. No promoter was identified matching these criteria. Rho-independent terminator structures were identified using the TransTermHP software tool [23]. Only rho-independent terminators at the correct genomic location with a score above 90 are displayed. Definition of the score is described in [23]. The program MEME was used for identification of conserved intergenic motifs in phage JG004 [24]. Acknowledgements The authors thank Gerd Doering, Burkhard Tuemmler and Michael Hogardt for providing clinical P. aeruginosa strains. Richard Muench helped with the TransTermHP analysis. We thank Dr. Elizabeth Murphy for proofreading. JG was supported by the DFG-European Graduate College 653. Electronic supplementary material Additional

file 1: Alectinib manufacturer Supplementary Table S1 and S2. S1: Genes of phage JG004 and their predicted function. S1: Predicted position of putative phage promoter. (PDF 191 KB) Additional file 2: Supplementary Figures. Contains Supplementary Figures S1 to S5. (PDF 225 KB) References 1. Strateva T, Yordanov D: Pseudomonas aeruginosa – a phenomenon of bacterial resistance. J Med Microbiol 2009, 58:1133–1148.PubMedCrossRef 2. Livermore DM: Has the era of untreatable infections arrived? J Antimicrob Chemother 2009,64(Suppl 1):i29–36.PubMedCrossRef 3. Skurnik M, Strauch E: Phage therapy: facts and fiction. Int J Med Microbiol 2006, 296:5–14.PubMedCrossRef 4. Summers WC: Bacteriophage therapy. Annu Rev Microbiol 2001, 55:437–451.PubMedCrossRef 5.