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: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: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:VarS/A is one of the global factors regulating diverse aspects of metabolism and virulence of bacteria including pathogenic Vibrio spp. An experiment to identify VarS/A-regulon in V. vulnificus revealed that a putative LuxR-type transcriptional regulator was down-regulated in ΔvarA mutant. To investigate the roles of this regulatory cascade from VarS/A to a LuxR-type regulator in V. vulnificus, the target gene regulated by a LuxR-regulator was identified and its expression was characterized.
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:The profiles of transcripts from the planktonic cells and biofilm cells of V. vulnificus were compared by using a V. vulnificus whole-genome microarray
Project description:IscR is a novel global regulator potentially contributing to the overall success in survival and pathogenesis of V. vulnificus by coordinating the regulation of various virulence factors. The profiles of transcripts from the V. vulnificus iscR mutant and the parental wild type were compared by using a V. vulnificus whole-genome microarray.
Project description:Dual transcriptome analysis of the V. vulnificus-infected human cells; To understand toxin-stimulated host-pathogen interactions, we set up dual transcriptome sequencing experiments using the human epithelial (HT-29) or immune (differentiated THP-1; dTHP-1) cells infected with the sepsis-causing pathogen Vibrio vulnificus, either in wild-type or the MARTX toxin-deficient backgrounds.