Below is an annotated figure showing the map of the phrGFP II-1 plasmid: Use the

Question

Below is an annotated figure showing the map of the phrGFP II-1 plasmid: Use the above figure to answer the following questions: What is the importance of having antibiotic resistance? Where on this plasmid is the antibiotic resistance located What is the promoter for the GFP sequence? What does MCS stand for and why are there two of these on the plasmid? Why are they useful? How are restriction enzymes important in plasmid engineering? Name 3 restriction sites on the phrGFP II-1 plasmid. What are the key differences between electroporation and chemical transformation?

Explanation / Answer

Q1. Antibiotic resistance helps in the selection of a transformed cell against untransformed cells with the plasmid. This plasmid would contain the cloned gene of interest.

Q2. pCMV is the promoter of GFP. CMV stands for cytomegalovirus.

Q3. MCS stands for multicloning site. It contains a set of different restriction sites which can be used for cloning the gene of interest. There are two MCS on each end of the GFP sequence.

Q4. Restriction enzymes help in cutting the DNA and inserting a desired piece of foreign DNA to make a recombinant molecule. The 3 sites are BamHI, HindIII and KpnI

Q5. Electroporation involves the use of electric pulse to create pores on the plama membrane and insert the recombinant plasmid. On the other hand chemical transformation makes use of chemicals such as Calcium chloride to make the plasma membrane porous enough for the recombinant plasmid to enter the cell.

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