Problem_Set3, bimm 101

Problem_Set3, bimm 101 - BIMM100 Fall 2008 Problem Set 3 1....

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BIMM100 Fall 2008 Problem Set 3 1. You would like to clone your favorite gene, Pancake (with a Flag tag), into a vector. The Pancake region in the insert diagram below contains both the promoter and protein coding DNA. a. Before your cloning begins, you need to choose a restriction enzyme to digest the insert and vector. NotI and PvuII both produce sticky ends (but at different DNA sequences) and SmaI produces blunt ends. Which enzyme do you choose and why? b. True or False –Ligation of the vector and insert can occur without treating the vector with a phosphatase. Explain. c. Following the ligation step, you need to transform your plasmid into bacteria. Why? What is one way to transform the bacteria with the plasmid?
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d. Once you’ve created the plasmid, you decide that you would like to purify your Pancake protein in bacteria. In order to do so you’ll need to modify your new plasmid. Name a modification you’ll need to make in the plasmid in order to express your protein in bacteria.
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This note was uploaded on 01/10/2009 for the course BIMM bimm 100 taught by Professor Dr.sato during the Fall '08 term at UCSD.

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Problem_Set3, bimm 101 - BIMM100 Fall 2008 Problem Set 3 1....

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