You will explore the use of Green Fluorescent Proteins (GFP), their structure, reactivity, purification, and identification. Green Fluorescent Proteins were first found naturally occurring in jellyfish by Japanese scientists decades ago as the compound that gave the jellyfish their bioluminescence. Since the discovery, scientists have isolated the protein and used it to see inside the cell of whatever they are studying easily. Just shining ultraviolet light on a cell containing GFP makes it light up green. There have since been many uses for GFP in biological research, including other derivatives of the protein that emit different wavelengths of light.
- View this brief introduction to GFP (you can focus just on the first 2 minutes as the rest get into some more details of GFP derivatives).
- Watch this video showing chromatography being used to purify bacteria and collect the GFP protein.
INTRODUCTION
In a Word document, first create a separate Title Page that contains the name of the project, along with your name and the date. Then, answer the questions below:
- The chromophore of GFP is what gives off the bright light. In the structure shown below, circle 1 amide, 1 amine, 1 carboxylic acid, and 1 alcohol.
- In the structure shown above, (a) how many amides are there? (b) amines? (c) carboxylic acids (d) and phenols?
- Referring back to the Project from Module 3, what IR bands would you expect to be prominent in an IR spectrum of the chromophore shown above?
- How are the proteins sorted on the column showed in the second video?
- Compare and contrast the chromatography method described above for GFP to the ones you used and the one you wrote about in your project for Module 2. (100-200 words)
- Referring back to Module 1, circle on the molecule shown below the area of the molecule that is aromatic and has conjugation. NOTE: You can either print or draw the molecule and then show your work for this question. All drawings should be signed and dated.
