A restriction map of the pGLO plasmid is shown below, with positionsSample Page where restriction enzymes cut shown as parentheticals. This plasmid can be used for a variety of purposes, including “tagging” a protein of interest with Green Fluorescent Protein (GFP) so researchers can determine where it localizes in a cell. You decide to do this with Your Favorite Gene (YFG), also shown above. You use SalHI to clone YFG in-frame with GFP a. Other than SalHI, what other enzyme do you need to do this 

b. To verify that cloning worked, you run restriction digests with three enzymes on both unmodified pGLO and with your recombinant plasmid. Draw the bands you would expect to see under the conditions listed above the gel (6 “lanes” total) The “AraC” feature in the map is the Arabinose operon. The expression of this operon is induced by the presence of Arabinose, and one of the proteins it produces is an Activator called AraBAD. AraBAD turns on the expression of GFP via the AraBAD operator. In another experiment, you have cloned another gene into the same plasmid, this time into the middle of the AraC operon, deactivating the function of AraBAD. You call this plasmid AraBAD-insert pGlo. c. Based on the description above, describe how much GFP would be expressed in cells where… No Arabinose + Arabinose No pGLO plasmids are present Unmodified pGLO is presen AraBAD-insert pGlo is present Both plasmids are present