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  • 1. A Comparison of Growth and Gene Expression in Two Species of Oysters By: Katie Fulkerson Advisor: Steven Roberts By: Katie Fulkerson Advisor: Steven Roberts

2. Oyster History

  • Oyster trade began in America in 1608
  • Habitat degradation and overharvest lead to population declines
  • Oyster culture seen as solution to saving natural stocks while continuing to meet consumer demand

3. Cultivated Oysters

  • Atlantic oyster ( Crassostrea virginica )
  • Kumamoto oyster ( Crassostrea sikamea )
  • European flat oyster ( Ostrea edulis )
  • Pacific oyster ( Crassostrea gigas )
  • Olympia oyster ( Ostrea conchaphila )

4. Pacific oyster(Crassostrea gigas) :

  • Valuable commercial species
  • Imported from Japan
  • Growth tends to be rapid when they are young and typically decreases when they reach 4-5 years of age.

5. Olympia oyster(Ostrea conchaphila):

  • Native species
  • This species experiences a slower growth rate, taking 4-5 years to reach market size (50mm).

6. Growth is dependent on

  • Genetics
  • Environmental factors
    • Water temperature
    • Food availability
    • Placement in the water column
    • Sediment type
    • Density of the bed

7. Objectives

  • Compare growth rates in two species of oysters from the same environment.
  • Identify genes that are likely involved in growth in Pacific and Olympia oysters.
  • Compare gene expression patterns from Pacific and Olympia tissues extracted during two periods of development.

8.

  • Oysters were grown in Agate Pass in Kitsap County, WA.
  • Measurements taken from umbo to edge, once a month from August to December 2007.
  • Tissues samples from the mantle, gills, and muscle were taken in September and November.

Objective 1: Growth Rates 9. 10. Objective 2: Gene Identification

  • Expressed sequence tags using bioinformatic techniques (KSPI, PKCIP, INSIG-2, P450)
  • One previously described gene (mGDF)

11. Molluscan growth differential factor (mGDF)

  • associated with the transforming growth factor beta family of proteins.
  • detected in the muscle ofO. conchaphilaand all tissues fromC. gigas .

m g ma L C. gigas m g ma O. conchaphila L 12. Kazal-type serine peptidase inhibitor domain 1 (KSPI)

  • A Serine proteinase inhibitor found in blood plasma, saliva, secretions of pancreas, seminal vesicles, and submandibular glands.
  • detected in all tissues samples of bothO.conchaphilaandC . gigas.

m g ma C. gigas L m g ma O. conchaphila L 13. Protein kinase C inhibitor protein 1 (PKCIP)

  • part of a family of conserved regulator proteins
  • plays regulatory roles in multiple cellular processes including differentiation, cell growth, secretion, and muscle contraction.
  • detected in the gill and muscle ofO. conchaphilaand all tissues ofC.gigas .

m g ma C. gigas L m g ma O. conchaphila L 14. Insulin-induced gene 2 protein (INSIG-2)

  • involved in metabolic activity, gene transcription, and cell growth.
  • not detected in any tissue of either species.

m g ma C. gigas L m g ma O. conchaphila L 15. Cytochrome P450 17-hydroxylase/lyase (P450)

  • a monooxygenase enzyme
  • functions in the metabolism of endogenous compounds such as aromatic hydrocarbons
  • In mollusks expression is highest in the digestive gland, but it is also found in blood cells, gills, foot, and gonads.
  • detected in the gill and muscle ofC.gigas .

m g ma C. gigas L m g ma O. conchaphila L 16. Objective 3: Gene Expression

  • Quantitative RT-PCR methods measured and compared gene expression levels in oyster tissue samples.

x x muscle muscle muscle Olympia Gill X mantle mantle mantle Pacific P450 INSIG2 PKCIP KSPI mGDF Species Tissues Sampled 17. 18. 19. 20. 21. Conclusion

  • Four genes associated with growth were successfully identified in these oysters.
  • Characterization of tissue expression patterns
  • Quantification of expression level in relation to different periods of development.
  • With the exception of KSPI all the genes examined varied in expression levels over time. This may indicate a switch in metabolic processes from the growing season (summer) to fasting season (winter).

22. Acknowledgements

  • Steven Roberts
  • Sam White
  • Vivianne Barry and Debbie Kay
  • Ken Fulkerson

23. ??Questions??