Biomedical Engineering: Bridging Medicine and Technology 1st Edition by Veronique Tran,Mark Saltzman

Exercise 20

Figure 3.31 shows the plasmid pBR322, which is commercially available from Invitrogen. The plasmid is 4363 bases in size; the sequence of bases within the plasmid is shown in the following table. Use only the information in this chapter to answer the following questions.
a. The restriction enzyme HindIII cuts this plasmid at only one site, at position 29. What sequence does HindIII recognize?
b. Identify a restriction enzyme that cuts the plasmid in only one location and cuts the gene for resistance to tetracycline (Tcr) but not the gene for resistance to ampicillin (Apr). What sequence does this enzyme recognize? What would be the value of using this restriction enzyme for cloning?
c. Use Figure 3.30 and the sequence below to find the amino acid sequence of the Tcr protein. Note the general location of the gene from the plasmid map. Also note that polypeptide sequences start with a start codon (AUG) and end with a termination codon (UAA, UGA, or UAG).