Comparative Transcriptomic Analysis of Salmonella in Desiccation
ABSTRACT: To explore the mechanisms of Salmonella desiccation resistance, we studied the transcriptomic responses in Salmonella Tennessee (Tennessee), using S. Typhimurium LT2 (LT2), a strain weakly resistant to desiccation, as a reference strain. In response to 2 h air-drying at 11% equilibrated relative humidity, approximately one-fourth of the ORFs in the Tennessee genome and one-fifth in LT2 were differentially expressed (> 2-fold). Among all differentially expressed functional groups (>5-fold) in both strains, the expression fold change associated with fatty acid metabolism was the highest, and constituted 51 and 35% of the total expression fold change in Tennessee and LT2, respectively. Tennessee showed greater changes in expression of genes associated with stress response and envelope modification than LT2, while showing lesser changes in protein biosynthesis expression. Expression of flagella genes was significantly more inhibited in stationary phase cells of Tennessee than LT2 both before and after desiccation. Salmonella Typhimurium LT2 ATCC strain 19585 (LT2) and Tennessee strain K4643 (Tennessee) were grown in TSB at 37 ºC with shaking for 20 h. Cells (~8 log CFU) for each strain were added on the discs and either stored at -80ºC directly, as control samples; or kept at 11% ERH at 25°C for 2 h and then transferred into -80 ºC, as desiccation treated samples. Three independent experiments were carried out for each condition.
Project description:Investigation of gene expression level changes in Salmonella typhimurium LT2 TA100 upon exposure to C60, compared to unexposed controls. The mutations engineered into this strain make it susceptile to mutagenic compounds. The Salmonella typhimurium TA100 strain used in this study is further described in Pedersen P, Thomsen E, Stern RM. 1983. Detection by Replica Plating of False Revertant Colonies Induced in the Salmonella Mammalian Microsome Assay by Hexavalent Chromium. Environmental health perspectives 51: 227-230. A 4 x 72K array study using total RNA recovered from triplicate cultures of Salmonella typhimurium LT2 TA100 exposed to C60 and triplicate cultures of controls that were not exposed to C60. Each 72K array measures the expression level of 4,504 genes from Salmonella typhimurium LT2 with seven 45 to 60-mer probe pairs per gene.
Project description:FabR ChIP-chip on Salmonella enterica subsp. enterica serovar Typhimurium SL1344 using anti-Myc antibody against strain with chromosomally 9Myc-tagged FabR (IP samples) and wildtype strain (mock IP samples) Overall design: IP sample (using anti-Myc antibody against Salmonella Typhimurium SL1344 strain encoding chromosomally 9Myc-tagged FabR) and control mock IP sample (using anti-Myc antibody against Salmonella Typhimurium SL1344 wildtype strain) were labeled with Cy5 and hybridized against a common genomic DNA reference, labeled with Cy3, on 2 S. Typhimurium LT2 whole genome tiling arrays
Project description:Obacunone is a limonoids present in Citrus species. We previously reported that obacunone was inhibitory to the cell-cell signaling in Vibrio harveyi and Escherichia coli O157:H7. In the present work we evaluated the effect of obacunone on the food borne pathogen Salmonella Typhimurium LT2 using cDNA microarray. The results demonstrate that obacunone exerts an antivirulence effect on S. Typhimurium LT2 by repressing SPI1 and SPI2. Furthermore, the effect of obacunone seems to be dependent upon EnvZ. One condition experiment, obacunone treated versus DMSO treated. Biological replicates: 3 control, 3 treatment, hybridized in dye-swapped design
Project description:Investigation of whole genome gene expression level changes in a Salmonella enterica serovar Typhimurium UK1 delta-iacP mutant, compared to the wild-type strain. IacP is resoponsible for the secretion of virulence effector proteins via the type III secretion system, thereby contributing the virulence of S. Typhimurium. The mutants analyzed in this study are further described in Kim et al. 2011. Role of Salmonella Pathogenicity Island 1 Protein IacP in Salmonella enterica Serovar Typhimurium Pathogenesis. Infection and Immunity 79(4):1440-1450 (PMID 21263021). A chip study using total RNA recovered from two separate wild-type cultures of Salmonella enterica serovar Typhimurium UK1 and two separate cultures of a mutant strain, Salmonella enterica serovar Typhimurium UK1 delta-iacP. Each chip measures the expression level of 4,302 genes from Salmonella enterica serovar Typhimurium.
Project description:Glucose-lysine based Maillard reaction products (MRPs) are introduced to the human body via food ingestion or are synthesized in vivo. Depending on reaction conditions, they represent a mixture of compounds with diverse chemical structures and physiological properties. MRPs may also be a potential nutrient source for Salmonella, a major foodborne gastrointestinal pathogen. This study was designed to determine the growth and transcriptional responses of S. Typhimurium LT2 to MRPs generated at low water activity to simulate the reaction occurring during food processing and storage. Maillard reactions between N-α-acetyl-lysine and glucose were varied in time (4, 23, and 143 h) to generate sub-samples with prevailing Amadori compound, advanced glycation end products (AGEs), or melanoidines respectively. The addition of MRP up to 5 mg mL-1 to defined media with 0.4% glucose had no effect on S. Typhimurium LT2 growth. In media, where the MRP sub-samples (4 and 23 h) served as the only carbon source, MRP assimilation by S. Typhimurium LT2 was observed as secondary logarithmic growth phases (0.063 h-1 and 0.072 h-1) after the growth on glucose available in the MRP reaction mixtures (0.479 h-1). The MRPs sub-sample with the highest concentration of melanoidines (143 h) also supported S. Typhimurium LT2 growth (0.368 h-1). Of the three MRPs sub-samples, the Amadori compound was the preferred carbon source for S. Typhimurium LT2 as evidenced by its almost complete disappearance (98%). Decreases in AGEs (37%) and melanoidines (15%), when incubated with S. Typhimurium LT2, also occurred. Transcription profiles of the cells grown on the MRPs fractions revealed predominant up-regulation of genes associated with the functional groups of energy metabolism, fatty and phospholipid metabolism, cellular process, and regulatory functions, and general down-regulation of the genes in the groups of amino acid biosynthesis, protein synthesis and transcription, transport and binding proteins, and DNA metabolism. The carbon flow in tricarboxylic acid (TCA) cycle partitioned by the glyoxylate cycle appeared to play an essential role in the assimilation of glucose-lysine-derived MRPs. The high expression level of numerous genes encoding hypothetical proteins or proteins with unknown function suggested the presence of genes whose role in glucose-lysine MRPs catabolism in Salmonella remains to be determined. This study was designed to determine the growth and transcriptional responses of S. Typhimurium LT2 to MRPs generated at low water activity to simulate the reaction occurring during food processing and storage. Maillard reactions between N-α-acetyl-lysine and glucose were varied in time (4, 23, and 143 h) to generate sub-samples with prevailing Amadori compound, advanced glycation end products (AGEs), or melanoidines respectively. Total bacterial RNA was isolated as previously described and the RNA samples were then converted to fluorescently-labeled cDNA and hybridized to S. Typhimurium microarrays version 5 (NIAID's Pathogen Functional Genomics Resource Center). Real-time PCR and physiological experiments were performed to validate the microarray data. In all supplementary files channel A = Cy3, channel B = Cy5.