Assignment Detail

Assignment ID: FG132598830

1. We will verify that we purified GFP using western blotting with a specific GFP antibody. What other antibody could be have used to achieve the same conclusion?
2. Could we have used in-gel fluorescence to see if we have purified GFP? Describe what in-gel fluorescence is and what we would have had to do differently to use this technique.
3. How could we use the purified GFP to make a specific antibody against GFP?
4. Imagine that we need GFP of higher purity. Discuss ways we could improve the GFP purification. In your answer, explain the use of “TAP-tags”.
5. GFP is a relatively easy protein to express in bacteria, as it is soluble and non-toxic, even at high expression proteins. Not all proteins are so easy to work with. Discuss the difficulties and strategies to overcome these difficulties when using bacteria to express:
i. proteins which are toxic to the bacteria
ii. membrane bound proteins