Assignment 3 - 2017

Vector features

This assignment must be submitted via UR Courses

One goal of this project is to subclone GeneA from the pKC7 plasmid (pBR322 plasmid with GeneA inserted) into the pUC19 vector. This assignment will help you understand why we might want to move an already cloned DNA fragment from one plasmid to another.

  1. pKC7 is a derivative of the pBR322 vector. Gene A was inserted into pBR322 using the restriction enzymes BamHI and HindIII. pBR322 is resistant to both tetracycline and ampicillin but pKC7 is only resistant to ampicillin. Explain why pKC7 is no longer tetracycline resistant. Hint: look carefully at the plasmid map for pBR322. (One to two sentences)
  2. The pUC18 vector is a derivative of The pBR322 DNA sequence was changed to create pUC18. Any change in a plasmid's sequence will result in a new plasmid that is a derivative of the starting plasmid; however, when we use this term to refer to vector plasmids, we generally mean that the changes made gave rise to a new vector that has desirable features the original vector did not. pBR322. How does the pUC18 vector differ from the pBR322 vector? Note that pUC18 and pUC19 are essentially the same vector so anything you find on pUC19 is true for pUC18.
  3. List three pUC18 features that make it a more popular vector than pBR322. For each feature, in one sentence, explain why that feature is desirable.

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