You are studying a human mRNA that you believe is controlled at the level of translation. You think that the 5’-UTR plays a role by binding to a protein that prevents the initiation of translation. To definitively determine the role of the 5’-UTR, you decide to clone the 5’UTR of the gene upstream of GFP open reading frame in a mammalian expression vector.

1. Describe in detail the experiments that you would perform using this vector to prove that the 5’ UTR plays a role in the control of translation.

2. Explain or draw what the result of the experiment would be if your initial hypothesis is correct.

3. What would be the possible effect of a mutation in the 5’-UTR that prevents that protein from binding?