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

3. Vibrio tubiashii

5. How do changes in the environment
influence Vibrio tubiashii physiology?

6. How do changes in the environment
influence Vibrio tubiashii physiology?
Information can be very informative
for other species

7. Parameters

8. Parameters

9. Experiments
oysters
pH gradient
control

10. Vibrio tubiashii
RE98

11. How do we assess alterations in
Vibrio tubiashii physiology?

12. How do we assess alterations in
Vibrio tubiashii physiology?
changes in expression of genes and proteins

13. How do we assess alterations in
Vibrio tubiashii physiology?
changes in expression of genes and proteins
basic biological information complementing
experimental and environmental studies

14. 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

15. Approach - Gene Expression
RNA Extraction
Select Genes
chitinase
vthB
ftsZ
serine protease
ompW qPCR
FliM

16. Approach - Gene Expression
RNA Extraction
Select Genes
chitinase
vthB
ftsZ
serine protease
ompW
FliM

17. Approach - Protein Expression
Protein Extraction
2D gels
ID differences

18. Experiments - gene expression
chitinase
oysters
control
Time (hrs)

19. Experiments - gene expression
vthB
oysters
control
Time (hrs)

20. Experiments - gene expression
pH gradient
chitinase vthB

21. Experiments - protein expression
oysters
control

24. Experiments - protein expression
oysters
control

25. pI MW
3.7 29k
4.5 49k
4.8 26k

26. pI MW
3.7 29k
4.5 49k
4.8 26k

27. 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)

28. 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