Project description:V. vulnificus is a marine bacteria that causes diseases in both mammals and fish. In both hosts, the iron concentration represents a key factor that greatly influences the virulence of this bacterium. To further define the gene repertoire that is regulated by iron concentration and Fur protein (the main transcriptional regulator in response to iron concentration) in V. vulnificus, we obtained a mutant in Fur and used DNA microarray technology to monitor the expression of the entire gene repertoire in response to iron. Global transcriptomic response was reconstructed by comparing the transcriptional profiles of the wild-type (R99) and Fur mutant strains in poor and rich iron conditions.
Project description:V. vulnificus is a marine bacteria that causes diseases in both mammals and fish. In both hosts, the iron concentration represents a key factor that greatly influences the virulence of this bacterium. To further define the gene repertoire that is regulated by iron concentration and Fur protein (the main transcriptional regulator in response to iron concentration) in V. vulnificus, we obtained a mutant in Fur and used DNA microarray technology to monitor the expression of the entire gene repertoire in response to iron. Global transcriptomic response was reconstructed by comparing the transcriptional profiles of the wild-type (R99) and Fur mutant strains in poor and rich iron conditions. To identify the genes that were under control of Fur, we compared the transcriptomic profile of the wild-type strain with the profile of a mutant strain in Fur protein; in contrast, to identify the genes that were under control of iron, we compared the transcriptomic profile of the wild-type strain grown in iron-rich conditions with the profile of the wild-type strain grown in iron-restricted conditions. For each one of the four samples, three replicates were performed, and RNA was sampled in the mid-log phase of growth (wild-type, 6h; Fur mutant, 6h; wild-type+iron, 5h; wild-type-iron,9h).
Project description:Vibrio vulnificus multiply rapidly in host tissues under iron overloaded conditions. To understand the effects of iron in the physiology of this pathogen we performed a genome-wide transcriptional analysis of this bacterium growing under three different iron concentrations. V.vulnificus CMCP6 cells were grown under three different iron concentrations (TSBS + EDDA 50uM, TSBS and TSBS + FAC 250 ug/ml) and samples taken at log phase. Keywords: Response to the iron concentration of the media
Project description:Vibrio vulnificus multiply rapidly in host tissues under iron overloaded conditions. To understand the effects of iron in the physiology of this pathogen we performed a genome-wide transcriptional analysis of this bacterium growing under three different iron concentrations. V.vulnificus CMCP6 cells were grown under three different iron concentrations (TSBS + EDDA 50uM, TSBS and TSBS + FAC 250 ug/ml) and samples taken at log phase. Keywords: Response to the iron concentration of the media Strains were grown to an OD600nm of 0.6 to 0.8 in TSBS, TSBS with the addition of 250 μg/ml FAC or TSBS with the addition of 50 μM EDDA. Three independent cultures of the V. vulnificus cells grown in each media, were combined and treated as a single sample for the RNA extraction to minimize culture variation. Two samples per condition were used for the microarray analysis. Cells were centrifuged and the pellets resuspended in RNAWiz reagent (Ambion®, Austin, TX). Total RNA was extracted from each strain according to the manufacturerâ??s instructions
Project description:Vibriosis caused by Vibrio vulnificus on eels represents an important threat for this specie under culture conditions. Development of new transcriptomic tools is essential to increase the knowledge of eel biology, that nowadays is scarcer. Therefore, using previous results obtained by 454 sequencing of the eel immune-enriched transcriptome, an eel-specific custom microarray have been designed. Gills transcriptomic pattern were analyzed as a principal portal of entry for pathogens in fish after 1h of bath infection with Vibrio vulnificus to describe gill immune response. Moreover, two different strains were used, vibro vulnificus wild type (R99) and rtx double mutant (CT285), to asses the virulence of these pathogen caused by MARTX.
Project description:Vibriosis caused by Vibrio vulnificus on eels represents an important threat for this specie under culture conditions. Development of new transcriptomic tools is essential to increase the knowledge of eel biology, that nowadays is scarcer. Therefore, using previous results obtained by 454 sequencing of the eel immune-enriched transcriptome, an eel-specific custom microarray have been designed. Gills transcriptomic pattern were analyzed as a principal portal of entry for pathogens in fish after 1h of bath infection with Vibrio vulnificus to describe gill immune response. Moreover, two different strains were used, vibro vulnificus wild type (R99) and rtx double mutant (CT285), to asses the virulence of these pathogen caused by MARTX. Adult european eels were bath infected with two Vibrio vulnificus strains, the wild type and double Rtx mutant (CT285). After 0, 3, 12h post-infection eel gills were sampled. Three individuals per experimental point were sampled, including a Control group and a Handling control group. Obtaining a total of 24 samples. The transcriptomic profile was described for each individual sample.
Project description:In order to analyze the transcripts of Arabidopsis thaliana (Col-0) and Vibrio vulnificus MO6-24/O simultaneously, Vibrio vulnificus MO6-24/O was infiltrated onto Arabidopsis leaves and then leaves were harvested at 0, 3, 6, 12, 24 and 48 h post-infiltration. A total of 31, 128, 303, 219 and 130 differentially expressed genes (DEGs) of Vibrio were up- and down-regulated at 3, 6, 12, 24 and 48 h post-infiltration (hpi). Meanwhile, differentially expressed genes (DEGs) were monitored at 3, 6, 12, 24 and 48 h post-infiltration. A total of 2,097, 1,839, 1,220, 1,170 and 1,383 genes were characterized at each time points in Arabidopsis. Our data clearly indicate that total transcripts of the marine bacterial pathogen V. vulnificus MO6-24/O are detected and analyzed in plant Arabidopsis and two organisms were inter-communicated at the same time under favorable conditions.
Project description:Stressosomes are stress-sensing protein complexes widely conserved among bacteria. Although a role in the regulation of the general stress response is well documented in Gram-positive bacteria, the activating signals are still unclear, and little is known about the physiological function of stressosomes in the Gram-negative bacteria. Here we investigated the stressosome of the Gram-negative, marine pathogen Vibrio vulnificus. We demonstrate that it senses oxygen and identified its role in modulating iron-metabolism.
Project description:Vibrio vulnificus (V. vulnificus) is an opportunistic human pathogen known for causing various illnesses such as gastroenteritis, skin and muscle necrosis, septic shock, and sepsis. This halophilic estuarine bacterium's growth and infection process involves adaptation to both the natural briny environments and the host. OmpR, a response regulator in the EnvZ/OmpR two-component regulatory system (TCS), is crucial for environmental adaptation and pathogenicity. This study focused on investigating the impact of OmpR in V. vulnificus by creating an ompR knockout strain (ΔompR). The ΔompR strain exhibited reduced tolerance to alkaline stress, shorter flagella, and decreased virulence in epithelial cell and mouse models compared to the wild-type (WT) V. vulnificus. RNAseq analysis revealed the downregulation of genes involved in metabolism, flagellum-dependent motility, and transcription factors in the ΔompR strain. OmpR was found to repress the expression of aphB in alkaline conditions, impacting the acid resistance system CadBA, while also positively regulating the transcription of various flagellar genes. These findings suggest that OmpR acts as a global regulator, orchestrating the expression of multiple genes in response to different environments and during host invasion.