PCSGA_vibrio

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PCSGA_vibrio

  1. 1. Characterizing the response of Vibrio tubiashii to changes in environment Tatyana Marushchak, Christin McLemore, Sam White Steven Roberts, Ralph Elston School of Aquatic and Fishery Sciences | University of Washington
  2. 2. Vibrio tubiashii
  3. 3. How do changes in the environment influence Vibrio tubiashii physiology?
  4. 4. How do changes in the environment influence Vibrio tubiashii physiology? Information can be very informative for other species
  5. 5. Parameters
  6. 6. Parameters
  7. 7. Experiments oysters pH gradient control
  8. 8. Vibrio tubiashii RE98
  9. 9. How do we assess alterations in Vibrio tubiashii physiology?
  10. 10. How do we assess alterations in Vibrio tubiashii physiology? changes in expression of genes and proteins
  11. 11. How do we assess alterations in Vibrio tubiashii physiology? changes in expression of genes and proteins basic biological information complementing experimental and environmental studies
  12. 12. How do we assess alterations in Vibrio tubiashii physiology? RATIONALE? changes in expression of genes and proteins basic biological information complementing experimental and environmental studies
  13. 13. Approach - Gene Expression RNA Extraction Select Genes chitinase vthB ftsZ serine protease ompW qPCR FliM
  14. 14. Approach - Gene Expression RNA Extraction Select Genes chitinase vthB ftsZ serine protease ompW FliM
  15. 15. Approach - Protein Expression Protein Extraction 2D gels ID differences
  16. 16. Experiments - gene expression chitinase oysters control Time (hrs)
  17. 17. Experiments - gene expression vthB oysters control Time (hrs)
  18. 18. Experiments - gene expression pH gradient chitinase vthB
  19. 19. Experiments - protein expression oysters control
  20. 20. Experiments - protein expression oysters control
  21. 21. pI MW 3.7 29k 4.5 49k 4.8 26k
  22. 22. pI MW 3.7 29k 4.5 49k 4.8 26k
  23. 23. Summary • Chitinase gene expression upregulated when adult oysters are present • Vthb (upstream of hemolysin) expression is elevated at 2hrs, then falls. • Changes in pH alter expression patterns • Several virulence associated pathways identified via protein expression • polar flagellin, lateral flagellin (motility), • OMP (survival & host invasion)
  24. 24. Future Direction • Expand the number of targets for gene expression • Examine other parameters • Coordinate with experimental and environmental sampling - relevance • Characterize the host response in relation to pathogen physiology

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