Pseudomonas aerugionsa PAO1 transcriptome cultured in MOPS Glycerol, MOPS Acetate
ABSTRACT: Drastic alterations in transcripts associated with central carbon metabolism and associated processes have been repeatedly observed a common expression program as P. aeruginosa adapts to the cystic fibrosis lung when compared to standard laboratory conditions. However, we have limited knowledge of how well these transcriptomic changes correlate with actual alterations in metabolic flux; the rate of turnover of molecules through metabolic pathways. We have compared the transcriptome of Pseudomonas aeruginosa PAO1 during growth on the carbon sources acetate and glycerol.
Project description:Pseudmonas aeruginosa PAO1 wild-type cultured in MOPS Glycerol compared to MOPS Glycerol Hypoxia (restricted oxygen). P. aeruginosa strain PAO1 was grown in 40 ml MOPS with glycerol as sole carbon source (triplicate), 37 °C with shaking (250 rpm) in baffled flasks (500 ml). For the restricted oxygen condition, P. aerugionosa was cultured in the same conditions, except non-baffled flasks were used, and the shaking speed was 80 rpm (gentle aggitation). A thin layer 10 ml of mineral oil was overlaid on top of these cultures to restrict oxygen transfer.
Project description:Anaplastic Lymphoma Kinase (ALK) is a tyrosine kinase receptor which is a clinical target of major interest in cancer, including neuroblastoma. To better understand ALK signaling, three different neuroblastoma cell lines (CLB-BAR, CLB-GE and SK-N-AS) were cultured for 1hr and 24hrs in control conditions or after treatment with the ALK inhibitors crizotinib or lorlatinib. RNA-Seq experiments were performed to determine the expression changes resulting from ALK inhibition. Together with parallel phosphoproteomic experiments, these data unveil several important conserved oncogenic pathways in neuroblastoma.
Project description:Objective: We analyzed changes in A. fumigatus gene expression profile at various stages of an in vitro model of aspergillosis to study the adaptation of A. fumigatus to the blood environment. Results: Most of virulence factors described to be involved in aspergillosis were not activated during the blood phase. We found three active processes to be activated in the later phase that may help to the adaptation: Iron homeostasis, a partial secondary metabolite cluster and the formation of detoxification enzymes. Conclusions: We propose that A. fumigatus is unable to grow in blood and it requires a metabolic change that allows the organism to shut down all uptake and energy-consume mechanisms, resulting in a resting mycelial stage. We performed gene expression profile by sequencing mRNA of A. fumigatus that were growm under two conditions, Minimal Medium (M) and human blood (B), and at different times: before placing the fungus in the final medium (pre), at 30' and at 180', with 2 biological replicates per condition.
Project description:For RNA sequencing, 3 days seedlings were moved to agar plates supplemented with 1/4 B&D media and susceptible zone of 14 days-old plants was harvested after specific treatment with water (Mock treated) or M.loti Nod factor 10-8M (NF). The total RNA was isolated from the susceptible zone (15 mm root pieces) using Nucleo spin RNA plant (Macherey-Nagel). Total RNA (> 0.8 g) from two biological replicas per sample was used by GATC Biotech (Germany) to prepare random primed cDNA library and for sequencing with Illumina HiSeq: read length 1 x 50bp. Gifu- L. japonicus wild-type, 4820- nfre-1 allele of Nfre, 38534- nfre-2 allele of Nfre.
Project description:We report that a transcriptional regulator that originated in the lineage that gave rise to multiple host-associated Candida species is a key component of the circuitry that governs the C. albicans cell surface composition. Specifically, we show that the transcription regulator ZCF21 controls the expression of genes encoding multiple cell surface proteins and cell wall modifying enzymes. Transcriptome (RNA-seq) analysis of 2 Candida albicans strains (reference strain and zcf21 deletion mutant) grown under 4 culture conditions (YPD broth at 30°C for 24h; YPD broth at 30°C in the presence of 15 mM caffeine for 24h; Todd-Hewitt agar at 37°C for 24h; Todd-Hewitt agar at 37°C for 24h in the presence of 5 mM caffeine).
Project description:New1 is not an essential gene but its deletion shows a cold-sensitive phenotype in yeast Saccharomyces cerevisiae. In this study, we compare the NEW1 knockout effect on translation using Ribo-Seq and RNA-Seq analyses.
Project description:Ribosome profiling (Ribo-Seq) and RNA-Seq analysis of eEF3 depletion in yeast (Saccharomyces cerevisiae). eEF3 depletion was induced by methionine in a modified strain where the native promoter was replaced by methionine repressible MET25 promoter. Conditional depletion enables us to study global effects of an essential gene.
Project description:Loss of New1 leads to a cold-sensitive phenotype of yeast Saccharomyces cerevisiae. In this study we investigated the effect of NEW1 knockout on translation using Ribo-Seq and RNA-Seq analyses.
Project description:Oligotropha carboxidovorans strain OM5 is an aerobic carboxidotrophic bacterium and potentially a promising candidate for such processes. We here performed RNA-Seq analysis comparing cells of this organism grown heterotrophically with acetate or autotrophically with CO2, CO and H2 as carbon and energy source and found a variety of chromosomally and of native plasmid-encoded genes to be highly differentially expressed. For RNA extraction and RNA-Seq, respectively, six aliquots of the sample of each culture replicate were pooled to gain enough cell mass and RNA, accordingly, while four biological replicates were processed for the RNA-Seq analysis. Total RNA was isolated from four biological replicates using Zymo Quick-RNA Miniprep Plus kit with bead beating and DNase-treatment (Zymo Research, Freiburg, Germany). Initially RNA quality was checked by Trinean Xpose (Gentbrugge,Belgium) and Agilent RNA Nano 6000 kit on Agilent 2100 Bioanalyzer (Agilent Technologies, Böblingen, Germany). Samples contaminated with DNA were treated with DNase again, cleaned as described above and rechecked by Xpose and Agilent Bioanalyzer. Finally RNA was free of DNA with an RNA Integrity Number (RIN) > 9 and rRNA Ratio [23s / 16s] > 1.5. Ribo-Zero rRNA Removal Kit (Bacteria) from Illumina (San Diego, CA, USA) was used to remove the ribosomal RNA molecules from the isolated total RNA. Removal of rRNA was checked by Agilent RNA Pico 6000 kit on Agilent 2100 Bioanalyzer (Agilent Technologies, Böblingen, Germany). RNA was free of detectable rRNA. TruSeq Stranded mRNA Library Prep Kit from Illumina (San Diego, CA, USA) was used to prepare cDNA libraries. TruSeq stranded cDNAs were sequenced paired end on an Illumina HiSeq1500 system, rapid mode using 70 bp read length.