Project description:As part of a broader study to identify genes that contribute to fitness of the human pathobiont Streptococcus agalacitae (group B Streptococcus), we identified a GntR-class transcription factor, named mrvR, which contributes to bacterial persistence in human amniotic fluid and multipe virulence phenotypes. In order to understand the transcriptome of mrvR, whole-genome transcriptomic analysis was performed with wild type group B Streptococcus and an mrvR deletion mutant at three growth phases.
Project description:Streptococcus agalactiae, also known as Group B streptococcus, emerged in the 1960s as a leading cause of septicemia and meningitis in neonates. It is also an increasing cause of infections in adults with underlying diseases. To characterize regulatory elements in this species we performed a genome-wide transcription start site (TSS) profiling and whole-transcript sequencing. TSS were identified by using a differential RNA-seq strategy, based on selective Tobacco Acid Pyrophosphatase (TAP) treatment and adapter ligation, which differentiates primary transcripts and processed RNAs. The accuracy and sensitivity of TSS identification were increased by combining differential RNA-seq analyses under eight conditions corresponding to variations in growth conditions and genetic backgrounds. Whole-transcript sequencing used a two-step adaptor ligation-based directional RNA-seq protocol and was performed under two experimental conditions with triplicate experiments to assess variations in gene expression in response to an acid stress
Project description:Transcriptome analysis of Streptococcus agalactiae (group B Streptococcus) grown under control conditions or coincubated with serine hydroxamate to induce the bacterial stringent response
Project description:Streptococcus agalactiae (Lancefield’s group B Streptococcus, GBS) is a major bacterial species of genus Streptococcus and has medical and veterinary importance by affecting mainly humans (Maione et al., 2005; Johri et al., 2006), cattle (Keefe, 1997) and fish (Mian et al., 2009). The GBS is the most important pathogen for the Nile tilapia, a global commodity of the aquaculture sector, causing outbreaks of septicemia and meningoencephalitis (Hernández et al., 2009; Mian et al., 2009). This study aimed to evaluate the global abundancy of proteins among the main genotypes of GBS isolated from fish identified in Brazil using a label free shotgun liquid chromatography-ultra definition mass spectrometry (LC-UDMSE) approach and to compare the differential expression of proteins identified between isolates from fish and human.
Project description:Streptococcus agalactiae (Lancefield’s group B Streptococcus, GBS) is a major bacterial species of genus Streptococcus and has medical and veterinary importance by affecting mainly humans (Maione et al., 2005; Johri et al., 2006), cattle (Keefe, 1997) and fish (Mian et al., 2009). The GBS is the most important pathogen for the Nile tilapia, a global commodity of the aquaculture sector, causing outbreaks of septicemia and meningoencephalitis (Hernández et al., 2009; Mian et al., 2009).
Project description:Regulation of gene expression in response to variable and often adverse environmental conditions is an essential component of microbial pathogenesis. We identified the two-component regulatory system CiaRH in a screen for genes essential for the survival of Streptococcus agalactiae (Group B Streptococcus, GBS) on exposure to in vitro models of environmental stress. We constructed site-directed, non-polar deletion mutations in the regulator gene ciaR and compared the growth of CiaR mutant GBS to wild-type GBS under stressed conditions. CiaR mutant GBS are more sensitive than wild-type GBS to elevated temperature, low pH, chemical mutagens and ultraviolet light; the mutants are also more sensitive to cell-wall active antibiotics and antimicrobial peptides. CiaR mutant strains are markedly attenuated in a mouse model of GBS sepsis. To determine the genes regulated by CiaR that account for these defects, transcriptional profiling was performed using DNA microarray analysis, comparing wild-type GBS to CiaR mutant GBS under non-stressed conditions.
