1.The yeast two-hybrid system is apowerful molecular genetic method to identify a protein(s) thatinteracts with a known protein or protein domain. You have isolatedthe glucocorticoid receptor (GR) and have evidence that is amodular protein containing an activation domain, a DNA bindingdomain and a second ligand binding-activation domain. Furtheranalysis reveals that in the pituitary cells the GR is anchored inthe cytoplasm in the absence of its hormone ligand. This resultlead you to speculate that GR binds to an inhibitory protein in thecytoplasm of the pituitary cells. (i) Describe how a yeast-twohybrid analysis system could help you to identify the protein withwhich GR interacts. ii) How would you specifically identify thedomain in the GR that binds the inhibitor? (20%)
1.The yeast two-hybrid system is apowerful molecular genetic method to identify a protein(s) thatinteracts with a known protein or protein domain. You have isolatedthe glucocorticoid receptor (GR) and have evidence that is amodular protein containing an activation domain, a DNA bindingdomain and a second ligand binding-activation domain. Furtheranalysis reveals that in the pituitary cells the GR is anchored inthe cytoplasm in the absence of its hormone ligand. This resultlead you to speculate that GR binds to an inhibitory protein in thecytoplasm of the pituitary cells. (i) Describe how a yeast-twohybrid analysis system could help you to identify the protein withwhich GR interacts. ii) How would you specifically identify thedomain in the GR that binds the inhibitor? (20%)
