: Construction of the baeR deletion mutant. (A) A single crossover between pEX18Tc containing baeR upstream and downstream sequences joined by a kan r cassette and the ATCC 17978 chromosome. (B) Two mechanisms by which the plasmid can integrate into the chromosome are diagrammed. (C) The suicide plasmid was excised by 10% sucrose counter-selection and selection of the in-frame baeR deletion strain with kanamycin. (TIFF 719 KB) Additional file 4: Figure S4.: Shuttle vector pWH1266 and verification of pWH1266 introduction into different strains of Acinetobacter baumannii. (A) pWH1266. (B) pWH1266 with kanamycin cassette insertion. (C) baeR insertion into the XbaI/XhoI restriction sites in pWH1266.

(D) Successful baeR

gene fragment insertion into the kanamycin cassette was deduced based on a change in the PCR band size from 1375 bp to 983 bp. AB1027, AB1028, and AB1029 represent the baeR reconstituted Ibrutinib in vivo strain, the baeR-overexpressing strain, and the A. baumannii ATCC 17978 strain with pWH1266, respectively. (TIFF 2 MB) Additional file 5: Figure S5.: baeR gene expression in different A. baumannii strains as determined by reverse transcription polymerase chain reaction. No baeR expression could be observed in AB1026. AB1027 was the baeR-reconstituted strain derived from AB1026, which had a baeR expression level similar to that of the wild-type strain. AB1028 and AB1029 represent the baeR-overexpressing strain and A. baumannii ATCC 17978 with pWH1266, respectively. (TIFF 840 KB) References 1. Fournier PE, Vemurafenib ic50 Richet H: The epidemiology and control of Acinetobacter baumannii in health care facilities. Clin Infect Dis 2006,42(5):692–699.PubMedCrossRef 2. Perez F, Hujer AM, Hujer KM, Decker

BK, Rather PN, Bonomo RA: Global challenge of multidrug-resistant Acinetobacter baumannii . Antimicrob Agents Chemother 2007,51(10):3471–3484.PubMedCentralPubMedCrossRef 3. Mendes RE, Farrell DJ, Sader HS, Jones RN: Comprehensive assessment of tigecycline activity tested against a worldwide collection of Acinetobacter spp. (2005–2009). Diagn Microbiol Infect Dis 2010,68(3):307–311.PubMedCrossRef 4. Lauderdale TL, Clifford McDonald L, Shiau YR, Chen PC, Wang HY, Lai JF, FER Ho M: The status of antimicrobial resistance in Taiwan among gram-negative pathogens: the Taiwan surveillance of antimicrobial resistance (TSAR) program, 2000. Diagn Microbiol Infect Dis 2004,48(3):211–219.PubMedCrossRef 5. Gordon NC, Wareham DW: Multidrug-resistant Acinetobacter baumannii : mechanisms of virulence and resistance. Int J Antimicrob Agents 2010,35(3):219–226.PubMedCrossRef 6. Rose WE, Rybak MJ: Tigecycline: first of a new class of antimicrobial agents. Pharmacotherapy 2006,26(8):1099–1110.PubMedCrossRef 7. Peleg AY, Adams J, Paterson DL: Tigecycline Efflux as a Mechanism for Nonsusceptibility in Acinetobacter baumannii . Antimicrob Agents Chemother 2007,51(6):2065–2069.PubMedCentralPubMedCrossRef 8.