Project description:We work with the bacterium Yersinia enterocolitica, a gastrointestinal pathogen. In this study we characterized a bacterial mutant strain called SOR17. We compared the protein extracts of SOR17 with that of the parental strain JB580v. Bacteria were grown in triplicate for 5 h at 37°C, and following bacterial disruption by French press, proteins were fractionated into soluble or membrane proteins. We also compared total protein extracts of bacteria grown for 16 h at 27°C. Two-dimensional electrophoresis were performed and proteins were stained with Coomassie brilliant blue. Spots whose abundance changed more than 2 fold were excised and sent to mass spectrometry for identification by either MALDI-TOF PMF or LC-MS/MS.
Project description:RNA-sequencing was preformed from RNA isolated from bacteria infected with the bacteriophage. In order to reveal the phage-host interactions between φR1-37 and Yersinia enterocolitica throughout the phage infection cycle, both the transcriptomes were scrutinized during all the stages of infection.
Project description:Orogastral infection of mice with Yersinia enterocolitical leads to HIF-1 alpha activation.To elucidate whether this HIF-1 alpha activation also results in a HIF-1 dependent gene programming, the transcriptomes from Peyers Patches of uninfected and Yersinia enterocolitica infected mice were analyzed by means of of microarray analyses using Affymetrix GeneChip probe arrays (MG-U74Av2). In total, 288 genes were differentially regulated three day after infection in PP compared with the expression of uninfected control mice. Of these 288 genes, 217 were found to be differentially upregulated and from these, 14 genes ( 6.5% of all upregulated genes) are well described to be regulated via HIF-1. These data indicate that orogatral infection with Y. enterocolitica results in HIF-1 dependent gene programmning Experiment Overall Design: Per group five C57BL/6 mice were infected orogastrally with 500 Million Yersinia enterocolitica. 1 and 3 days after infection, Peyers Patches were removed and total RNA was prepared. In parallel, RNA was isolated from uninfected mice. The generation of fragmented cRNA was performed following the manufacturers instructions and used for hybridization onto GeneChip arrays MG-U74Avs2. Analysis of microarray data was performed using the Affymetrix Microarray Suite 5.0, Affymetrix Mining Tool 3.0. A median signal log2 ratio (SLR) grater than 1.5 or less than -1.5 was considered a significant change.