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Team:Edinburgh/Results/Glycogen2
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Glycogen Assay 2
Background and Aims
This assay was designed to determine qualitatively the effects of our glgC and glgC16 BioBricks on glycogen synthesis. We hypothesised that cells transformed with glgC or glgC16 BioBricks would produce more glycogen than control cells, since glgC would be overexpressed and freed from control by its usual promoter in transformed cells. glgC16 is a version of glgC which contains a mutation rendering it immune to negative feedback. Hence, we expected glgC16-transformed cells to produce even more glycogen than glgC-transformed cells. Finally, cells should produce more glycogen when grown in a glucose-rich medium.
Procedure (3~4 Oct 2008)
- A lac promoter was added to the glgC and glgC16 biobricks.
- JM109 E. coli cells were transformed with...
- glgC BioBrick + lac promoter.
- glgC16 BioBrick + lac promoter.
- (Control = not transformed)
- Cells were grown overnight in LB and LB+40mM glucose.
- Cells were resuspended in iodine reagent to test glycogen levels. Cells containing more glycogen would stain a darker brown than cells containing less glycogen.
Results
Results of the assay were as follows:
These results confirm that:
- glgC and glgC16 BioBricks resulted in enhanced glycogen production, especially when cells are grown in a glucose-rich medium.
- It is not obvious in the photo above, but cells with glgC16 stained darker than cells with unmutated glgC, indicating that the mutation increased glycogen production.
