No negative staining was performed. Linoleic acid survival assay S. aureus linoleic acid survival assays were performed essentially as described by Kenny et al. . Briefly, serial dilutions of overnight cultures (2.5 μl spots) were plated in duplicate onto BHI agar, pH 6.0, containing 0 mM or 1 mM linoleic acid. All agar media contained a final concentration of 1% ethanol. Colonies were counted after overnight incubation at 37°C. Mean values were compared using Student’s t test. S. saprophyticus survival assays were performed similarly, but with agar plates containing 5 mM linoleic acid, supplemented with 0.85 M NaCl. Structural predictions of SssF
Secondary structure and three-dimensional fold predictions were performed using PSI-PRED  and Phyre , respectively. Acknowledgements This work was supported by buy KPT-330 grants from the Australian National LXH254 chemical structure Health and Medical Research Council to M.A.S. (569676) and S.A.B. (511224), and a University of Queensland Early Career Researcher grant to S.A.B. M.A.S. is supported by an Australian Research Council (ARC) Future Fellowship (FT100100662) and S.A.B. is supported by an ARC Australian Research Fellowship (DP0881247). Electronic supplementary material Additional file 1: Table S1. Predicted protein-coding genes of pSSAP2. (DOC 156 KB) Additional file 2: Figure S1. ClustalW alignment of the C-terminal 402 amino acid residues of S. saprophyticus MS1146
SssF protein (61% of entire sequence) with corresponding sequence from other staphylococcal Lonafarnib in vitro SssF-like proteins, showing clusters of amino acid conservation. Only one representative protein from each www.selleckchem.com/products/JNJ-26481585.html species is shown. Sequences are sorted (in descending order) by similarity to S. saprophyticus MS1146 SssF sequence,
which ranges from 31.1% (S. pseudintermedius HKU10-03) to 48.5% (S. carnosus TM300). Jalview was used to colour-code the alignment by percentage identity. The C-terminal sortase anchor motifs are indicated by a red box. GenBank accessions for the SssF-like proteins are as follows: S. carnosus TM300, CAL29334; S. capitis SK14, EEE48467; S. caprae C87, EFS16450; S. epidermidis RP62A, AAW53125; S. warneri L37603, EEQ79103; S. haemolyticus JCSC1435, BAE03665; S. hominis SK119, EEK11979; S. aureus NCTC 8325, ABD31969; S. lugdunensis HKU09-01, ADC86449; S. pseudintermedius HKU10-03, ADV06726. (TIFF 4 MB) References 1. Schappert SM: Ambulatory care visits to physician offices, hospital outpatient departments, and emergency departments: United States, 1997. Vital Health Stat 1999,13(143):1–36. 2. Foxman B, Barlow R, D’Arcy H, Gillespie B, Sobel JD: Urinary tract infection: self reported incidence and associated costs. Ann Epidemiol 2000,10(8):509–515.PubMedCrossRef 3. Hooton TM, Stamm WE: Diagnosis and treatment of uncomplicated urinary tract infection. Infect Dis Clin North Am 1997,11(3):551–581.PubMedCrossRef 4.