RNA-seq analysis of Acinetobacter baumannii AB5075 and grey colony variants having a copy of ISAba13 in the K-locus
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ABSTRACT: RNA-seq was used to compare the transcriptomes of wild type A. baumannii AB5075 and a spontaneously arising grey colony variant that carries a copy of the insertion sequence ISAba13 within the gtr52 gene.
Project description:The experiment contains ChIP-seq data for Acinetobacter baumannii strain AB5075 encoding 3xFLAG tagged H-NS. Experiments were done with or without ectopic expression of the truncated H-NS-39 protein (corresponding to the H-NS multimerization surface). The strain was grown at 37 degrees in LB medium and crosslinked with 1 % (v/v) formaldehyde. After sonication, to break open cells and fragment DNA, immunoprecipitations were done using anti-FLAG antibodies against. Libraries were prepared using DNA remaining after immunoprecipitation.
Project description:The experiment contains 3C-seq data for Acinetobacter baumannii strain AB5075 with different genetic alterations. The strain was grown at 37 degrees in LB medium and nucleoprotein was cross-linked using formaldehyde. Genomic DNA was isolated and digested with NlaIII before being ligated with T4 ligase. Sequencing was then used to identify junctions between ligated DNA sequences.
Project description:The experiment contains native Tn-seq data for Acinetobacter baumannii strain AB5075 with different genetic alterations. The strain was grown at 37 degrees in LB medium and genomic DNA was isolated. We then used PCR to select for DNA regions containing a junction between ISAba13 and chromosomal DNA. Libraries were then prepared using these DNA fragments.
Project description:The experiment contains native Tn-seq data for Escherichia coli strain MG1655. The strain was grown at 37 degrees in LB medium and genomic DNA was isolated. We then used PCR to select for DNA regions containing a junction between insH3 and chromosomal DNA. Libraries were then prepared using these DNA fragments.
Project description:The nosocomial pathogen Acinetobacter baumannii is a frequent cause of hospital acquired infections worldwide, and a challenge for treatment due to its evolved resistance to antibiotics, including carbapenems. To gain insight on A. baumannii antibiotic resistance mechanisms, we analyzed the protein interaction network of a multidrug-resistant A. baumannii clinical strain Ab5075. Using in vivo chemical cross-linking and mass spectrometry, we identified 2,068 non-redundant cross-linked peptide pairs containing 245 intra- and 398 inter- molecular interactions. Outer membrane proteins OmpA and YiaD, and carbapenemase Oxa-23 are hubs of the identified interaction network. Eighteen novel interactors of Oxa-23 were identified. Interactions of Oxa-23 with outer membrane porins OmpA and CarO were verified with co-immunoprecipitation analysis. Furthermore, transposon mutagenesis of oxa-23 or interactors of Oxa-23 demonstrated changes in meropenem or imipenem sensitivity in Ab5075. These results provide the first view of a porin-localized toxin inactivation model and increase understanding of bacterial antibiotic resistance mechanisms.
Project description:Cappable-seq was used to map transcription start sites globally in Salmonella Typhimurium SL1344. In addition to naturally occurring plasmids pSLT and pCol1B9, the cells carry plasmid pAMNF. The latter can be used for low level constitutive expression of a transcription factor of interest, though in these experiments no such ectopic transcription factor expression was used.
Project description:Cappable-seq was used to map transcription start sites globally in Salmonella Typhimurium SL1344. In addition to naturally occurring plasmids pSLT and pCol1B9, the cells carry derivatives of plasmid pAMNF, or pAMNM, that drive low level constitutive expression of epitope tagged MarA, SoxS, Rob or RamA derivatives.
Project description:The dimerization and binding of DNA by BldD is affected by its interaction with cyclic di-GMP. The D116A mutant of BldD is partially impaired in its biding of cyclic di-GMP. ChIP-Seq was carried out to determine the difference in the degree of DNA binding by the wild type and D116A mutated BldD in Streptomyces venezuelae.
Project description:Analysis of the effect of human pleural fluid (HPF) or human serum albumin (HSA) on the transcriptome of Acinetobacter baumanii AB5075