Project description:Objectives: The Burkholderia pseudomallei bprR and bprS genes are predicted to encode the response regulator and sensor kinase components respectively of a two component signal transduction system. Inactivation of either single component of this two component signal transduction system led to attenuation of B. pseudomallei virulence and also reduced motility. However, inactivation of both components resulted in wild-tpe virulence and motility. Therefore, we used RNA-seq to compare the transcritptomes of the three mutant strains (bprR single, bprS single, bprRS double mutant) with the wild-type parent strain in order to identify the genes differenty regulated in the different mutants.
Project description:This study describes the global transcriptional changes upon deletion of a hypothetical gene BPSS1356. The BPSS1356 deletion strain of B. pseudomallei was generated via homologous recombination. Global gene expression profiling of wild type and ∆BPSS1356 mutant strains was performed to elucidate of biological functions of BPSS1356.
Project description:We report the transcriptome of Burkholderia pseudomallei type VI secretion regulator TctR mutant grown in rich media compared to wild type. The RNA-seq studies confirmed the role of TctR as a negative regulator of T6SS-2, a positive regulator of T6SS-6 and suggest a potential role in regulation of the T6SS-3 and T6SS-4 gene clusters.
Project description:Comparison of the transcriptomes of a Burkholeria pseudomallei bprR [bpss0688] mutant, a bprS [bpss0687] mutant and a bprRS double mutant with the wild-type B. pseudomallei K96243 parent strain.
Project description:We report the application of single-molecule-based sequencing technology for high-throughput profiling of DNA methylations in Burkholderia pseudomallei.
Project description:Burkholderia pseudomallei is a soil-dwelling bacterium which has to survive not only under harsh environmental conditions, but also within various hosts where it can cause the infectious disease melioidosis. The ability, to quickly adapt to these different conditions, is based on its huge genome which encodes for complex regulatory networks. Among them are more than 60 genes belonging to the group of LysR-type transcriptional regulators (LTTRs). Here we analyzed a B. pseudomallei mutant harboring a transposon in the gene BPSL0117 annotated as a LTTR, which we named gvmR (globally acting virulence and metabolism regulator). The gvmR mutant displayed a growth defect in minimal medium and macrophages in comparison with the wild type. Moreover, inactivation of GvmR rendered B. pseudomallei avirulent in mice indicating a critical role of GvmR in infection. These defects of the mutant were rescued by ectopic expression of gvmR. To identify genes whose expression is modulated by GvmR, global transcriptome analysis of the B. pseudomallei wild type and gvmR mutant was performed using whole genome tiling microarrays. Transcript levels of 190 and 142 genes were found to be up- and downregulated in the gvmR mutant relative to the wild type. Among the most downregulated genes in the gvmR mutant were important virulence factor genes (T3SS3, T6SS1 and T6SS2), which might provide an explanation for the virulence defect of the gvmR mutant. In addition, expression of genes related to amino acid synthesis, glyoxylate shunt, iron-sulfur cluster assambly and syrbactin metabolism (secondary metabolite) was decreased in the mutant. Furthermore, inactivation of GvmR increased expression of genes involved in pyruvate metabolism, ATP synthesis, malleobactin and porin genes. Quantitative RT-PCR verified the differential expression of 27 selected genes. In summary, our data show that GvmR acts as an activating and repressing global regulator that is required to coordinate expression of a diverse set of metabolic and virulence genes for survival in the animal host and under nutrient limitation.
