Chapter 9, Problem 15P

A recombinant DNA molecule is constructed using a plasmid vector called pMBG36 that is 4271 bp long. The pMBG36 plasmid contains a so-called polylinker that has a single site for each of several restriction enzymes, including BamHI (5′ G^GATCC 3′) and EcoRI (5′ G^AATTC 3′). The sequence of the polylinker region of pMBG36 is shown below; the dots indicate the large majority of the vector that is not shown. You now cut

pMBG36 with EcoRI and insert into it a fragment of the DNA previously shown in Problem 4 that is also cut with EcoRI.

a.	Write out as much of the DNA sequence of the resultant recombinant DNA molecule as is possible. Two answers are possible; you need to show only one.
b.	Why are there two possible answers to part (a)?
c.	How many recognition sites for BamHI will be found in the recombinant DNA molecule shown in your answer to part (a)?
d.	If you cut this recombinant DNA molecule with BamHI and run the digest on a gel, how many bands would you see and how large would they be?
e.	How many recognition sites for EcoRI will be found in the recombinant DNA molecule shown in your answer to part (a)?
f.	If you cut this recombinant DNA molecule with EcoRI and run the digest on a gel, how many bands would you see and how large would they be?