13 Jun What effect would this have if the binding site is involved in positive regulation
Question
TAKE-HOME EXTRA CREDIT
These questions are open-note, open-book, chat with all of your colleagues questions. I ask only a couple things:
1. Use your own words. I will pick up copy-and-paste, and it will make me cranky. You won’t like me cranky.
2. Cite any and all references you use. I don’t care about format, so long as it contains all of the relevant information. Questions? Email me.
Nicole and I will set up a drop-box on D2L for you to submit these answers by 5PM Wednesday, April 29. You don’t have to do all of them – you can pick and choose.
I will post these questions and all of the relevant information on D2L after the exam.
1. You have cloned a region of DNA that contains a BINDING SITE (and ONLY a binding site!) for a regulatory protein from a prokaryotic operon. You put this clone into a high copy plasmid and introduce it into a cell that is wild type for the regulation you are studying. (5 pts)
A) What effect would this have if the binding site is involved in positive regulation and why?
B) What effect would this have if the binding site is involved in negative regulation and why?
2. You have received two plasmids in the mail, pHS1 and pEC1. (5 pts)
pHS1 is a plasmid that contains a 7 kb EcoRI fragment from the human genome
pEC1 is a plasmid that contains a 7 kb EcoRI fragment for the E. coli genome
Unfortunately, the tube labels washed off in transit. Describe how you would determine which tube contains which fragment. Note that you want to tackle this without doing any DNA sequencing.
3. Describe the process of producing a cDNA library for genes ONLY expressed in one tissue of your eukaryotic organism of choice. Include all relevant enzymes. (10 pts)
4. Describe at least two DNA sequencing methods, including chemistry, positives, and minuses of each technique. (10 pts)
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