streptococcus pneumoniae pathogenesis sam king cmp and bcmm meeting
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Streptococcus pneumoniae pathogenesis
Sam KingCMP and BCMM meeting
![Page 2: Streptococcus pneumoniae pathogenesis Sam King CMP and BCMM meeting](https://reader035.vdocument.in/reader035/viewer/2022072016/56649ee75503460f94bf7c5e/html5/thumbnails/2.jpg)
Projects
• Does pneumococcal genomic variation contribute to development of different disease states
• Structure function analysis of pneumococcal transporters
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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?
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To Identify pneumococcal sequences that contribute to HUS
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Aims
1. Identify pneumococcal loci that potentially correlate with development of HUS
2. Identify pneumococcal HUS enriched sequences
3. Determine the biological contribution of these sequences to pathogenesis
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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
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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
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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
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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?
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Biological consequences of these HUS enriched sequences
• Correlation but not causation• Contribution to pathogenesis
– Bioinformatics– Biochemical assays– Genetic approaches
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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
ATP
ADP + Pi
ADP + Pi