This is the table I came up with but may not be what the plates are exactly show
ID: 227598 • Letter: T
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This is the table I came up with but may not be what the plates are exactly showing. A loose interpration needs to be made as I was absent and unable to view the plates. I only have the attached image. What I am stuck on are the questions. Just need a brief answer for them.
What do the glycerol and glucose do in this pGLO experiment?
**Please help with any information whatsoever for #2 and 3. Thank you!!
1. Prepare a table reporting your results
2. Based on your results, what conclusions can you draw concerning whether the glucose transporter or glycolytic flux is involved in inhibiting adenylate cyclase.
3. Suppose that you had made a second set of plates with the same compounds as before, but that you had added 1% cAMP to the media to each. How would the results have changed?
Table Agar Plates with: LB, ampicillin, and arabinose LB and ampicillin LB, ampicillin, arabinose, and glycerol LB, ampicillin, arabinose and glucose Glow under UV light: Yes No Yes Yes Growth on plates: Yes Yes Yes YesExplanation / Answer
If we put pGLO into the plates having the LB/amp and the plates having the LB/amp/ara, then only the plate labeled LB/amp/ara will glow. pGLO have the gene for green color or brightness under UV light. It have gene for Arabinose (ara) C protein. B-lactomase helps in the degradation of ampilcillin which makes the pGLO plasmid ampilcilin resistant. Arabinose removes the GFP after that RNA polymerase get activated which starts the expression of gene. It means that Transformation using the pGLO will glow only in those conditions when arabinose will present inside there.
Glucose acts as repressor where it inhibits the activity of adenylate cyclase. This will lower the cAMP concentration. In lack of cAMP the Catabolism activator protein (CAP) loses its affinity to bind with promoter of expression system to express the gene. Hence RNA polymerase will unable to start transcription. If you will add the 1% cAMP it will help in expression of gene but other factor are also required i.e. Arabinose/LB/amp.
In glucose Transport Model, Glucose regulates the expression of gene by inhibiting the adenylate cyclase. There is a factor EIIAglc which will not bind with adenylate cyclase because phosphate groups are absent. Non interaction will decrease the level of cAMP. Hence the expression will slow down. If glucose will not transport then the phosphate will not dissociate from EIIAglc factor. And further process of expression will continue.
In glycolytic flux model, the phosphate working in reverse manner in comparison to the glucose transport model. Glucose level decreases then Glycerol concentration will increases. These phosphorylated intermediate will prevent EIIAglc to activate adenylate cyclase. Hence the non active form will unable to start the expression process.
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