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Streptococcus pneumoniae pathogenesis

Streptococcus pneumoniae pathogenesis. Sam King CMP and BCMM meeting. Projects. Does pneumococcal genomic variation contribute to development of different disease states Structure function analysis of pneumococcal transporters . Pneumococcal diversity. Pneumococcal bacteremia

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Streptococcus pneumoniae pathogenesis

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  1. Streptococcus pneumoniaepathogenesis Sam King CMP and BCMM meeting

  2. Projects • Does pneumococcal genomic variation contribute to development of different disease states • Structure function analysis of pneumococcal transporters

  3. Pneumococcal diversity Pneumococcal bacteremia Hemolytic uremic syndrome (HUS) Is there something different about these isolates? Can we use that knowledge to develop tests to identify patients at risk or therapeutics?

  4. To Identify pneumococcal sequences that contribute to HUS

  5. Aims • Identify pneumococcal loci that potentially correlate with development of HUS • Identify pneumococcal HUS enriched sequences • Determine the biological contribution of these sequences to pathogenesis

  6. Identify pneumococcal loci that potentially correlate with development of HUS • Sequence six pneumococcal HUS isolates • Two methodologies • Next generation (Biomedical Genomics Core) • 30 million reads for each strain • Short (~100bp), high accuracy • Has to be aligned to a sequenced genome • Third generation sequencing (EA sequencing) • Longer reads (~3kb) • Accuracy low • Will allow generation of de novo sequence

  7. Analysis of sequence data • Biomedical Genomics Core (Post doctoral scientist) • Will generate accurate de novo sequence using info from both techniques • Compare sequence with 16 sequenced genomes • King lab • Prioritize sequence variants for analysis • Conserved in the majority of HUS isolates • Absent in the majority of non-HUS isolate • Introduce or remove open reading frames • Change promoter or coding sequence of predicted extracellular proteins

  8. Identify pneumococcal HUS enriched sequences • Screen presence of up to 50 variants • In up to: • 50 HUS isolates • 50 non-HUS blood isolates • Determine if there is a significant correlation with HUS

  9. Identify pneumococcal HUS enriched sequences • Will 50 strains be enough? • If pneumococcal HUS isolates are more closely related genetically how will that affect our data?Can we take account of that?

  10. Biological consequences of these HUS enriched sequences • Correlation but not causation • Contribution to pathogenesis • Bioinformatics • Biochemical assays • Genetic approaches

  11. Structure function analysis of pneumococcal transporters • ATP binding cassette transporters • Conserved protein family • Five components Substrate binding protein Permease proteins ATPase (nuclear binding domains) ATP ADP + Pi ADP + Pi ATP

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