PS_1 - Biology 115 Winter 2011 Problem Set#1 1 You want to...

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Biology 115, Winter 2011 Problem Set #1 1. You want to express the Purple gene in bacteria. This is a mouse gene responsible for bright purple streaks observed in the hair of some mouse species. Surprisingly it naturally lacks introns. Question A. A lab at UCSC previously cloned Purple gene with its natural promoter into a cloning vector . Describe two methods they could have used to obtain the Purple DNA sequence. Question B. You also have a bacterial expression vector . Restriction maps for both of these are shown below. (A restriction map is a cartoon/map showing the relative order and position of restriction sites in a piece of DNA). What are the functions of the labeled landmarks of these plasmids? Question C. Describe a strategy for cloning the Purple gene into the expression vector. This requires putting the Purple gene’s coding sequences -indicated by the boxed sequences-in the correct orientation downstream of the bacterial promoter. (Note: the process of creating a recombinant from previously cloned genes is often called subcloning ) 2. The recognition sequence for Xho I is 5'-CTCGAG-3' 3'-GAGCTC-5' and the recognition sequence for Sal I is 5'-GTCGAC-3' 3'-CAGCTG-5' Both of these restriction enzymes cut their sites so as to leave 4 basepair 5' overhangs . Question a.
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PS_1 - Biology 115 Winter 2011 Problem Set#1 1 You want to...

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