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Green Florescent Protein as a Marker for Gene Expression. Role of GFP in Aequorea victoria. Photoprotein aequorin stimulated by calcium ions Aequorin catalyzed to higher energy state: Apoaquorin 3 . Blue light released as Apoquorin relaxes to ground state.
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Role of GFP in Aequoreavictoria • Photoproteinaequorinstimulated by calcium ions • Aequorin catalyzed to higher energy state: Apoaquorin 3. Blue light released as Apoquorin relaxes to ground state
Role of GFP in Aequoreavictoria 4. Apoaequorin excites the chromophore in GFP 5. GFP produces green lightas it returns to ground state
Our Problem: • Can GFP be expressed without the coenzymes in A. victoria? • If so, can GFP be expressed in other organisms?
Experiment 1 – GFP in E. Coli Bacteria on the right side express GFP while under the effect of irradiation with UV light Photographs were taken during irradiation
Experiment 2 – Transformation of C. Elegans Spliced under control of the mec-7gene promoter The bright green centers are two touch receptors: ALMR, PLMR
Applications of GFP Transformation • Track any gene visually • More reliable than current techniques • Useful for clear bodied organisms(C. elegans)
Work Cited M Chalfie, Y Tu, G Euskirchen, WW Ward, and DC Prasher Science 11 February 1994: 263 (5148), 802-805.
Response Letter I really appreciate the input put forth by my peers on this piece. I realize how “rough” the rough draft was. This assignment was especially difficult because although I am a “scientist in training,” I’m not quite at the level to write so technically. However, the feedback I received definitely helped me to develop this project. Reviewer one mentioned that I could use a lot more in the document itself. I disagreed. I felt that this PowerPoint was supposed to touch on the topic and prove its uses. Members of the audience seriously interested in this would take the steps to find the paper and repeat the necessary steps for their own interested. I would like to thank this reviewer on the compliments for my design, though. Reviewer two enjoyed the sparse rhetorical questions to break the jargon. She also mentioned that I may want to remove the first two slides on how GFP works. I disagreed. There are a large variety of attendees at these conferences some are undergraduate interns and many only know about C. elegans. I thought it may be worthwhile to spend the extra 5 minutes getting everyone caught up. Reviewer three complimented me on the balance between the words on the slide and the script. However, she saw a lack of flow in the first slide and the script. I completely agreed. I even forgot to mention myself and credentials. I changed this section, moving from “welcome” to “thank you”, and then from my credentials and what I do to the research pertinent to the presentation. I also messed with the word balance on slides three and five so it was less like the presenter was reading from the slides. Again, I would like to thank my editors for any contributions they had. Anything and everything was considered while editing the draft.