Project description:We used transposon insertion sequencing (Tn-Seq) to identify the genes that are required for in vitro growth and intramacrophage growth of the live vaccine strain of F. tularensis (LVS).
Project description:The goal of this study is to determine the host response of human epithelial cells during infection with Francisella Tularensis. For this purpose, A549 human epithelial cell line was infected with Francisella tularensis spp. holarctica strain LVS for different times of infection, in duplicates. At different times post infection (0.5/1/3/6/12/24 hours post infection) cells were harvested and total RNA was extracted. RNA-seq libraries were constructed and sequencing of 100bp paired-end was performed on the Illumina NovaSeq 6000 system. Sequencing yielded about 22M reads per sample that were mapped to the human genome (Human: GRCh38) resulting with the identification of 21,066 transcripts. The expression of the infected samples was compared to mock sample, and RNA ratios were clustered using partitioning clustering. This approach allowed clustering of the cellular transcripts into 5 distinct classes based on similarities in temporal expression profiles. We next carried out GO term enrichment analysis for each of these five cluster. Our study represents the first detailed analysis of human epithelial response to Francisella tularensis infection, and provide a framework for comparative investigations of genes and mechanisms that may contribute to the infection.
Project description:Comparison of enriched membrane fractions of Francisella tularensis subsp. holarctica strain FSC200 and its DsbA mutant by SILAC analysis.
Project description:Three pairs of total RNA preparations from F. t. LVS grown under iron-replete and iron-restricted conditions were used in the DNA microarray. Seven arrays were used to examine the genes affected by iron limitation. Keywords: Dual Channel (Cy5/Cy3) arrays
