BIOL 5060 Lecture Notes - Lecture 17: Phosphodiester Bond, Electrophoretic Mobility Shift Assay, California Institute Of Technology
Document Summary
Protein-dna interactions: dnasei footprinting- will show region of protein binding site, ggatcc- palindrome; bahi site, test the sequence specificity of the interaction by making cold competing dna molecules, do a band shift , 1. Translational fusions- carry out emsa with an extract making wild-type protein versus an extract making a hybrid version of the protein. If it"s a functional fusion protein, it should shift: would prove that protein is part of the complex because fusion protein affected shifting. If the fusion protein loses binding activity no band: no shift would tell us that fusion protein affected binding activity protein is a. If fusion or wild-type protein isn"t involved in binding, there will be no shift: protein is not part of the complex (bands are unaffected by fusion and look them same, 2. Super-shift using antibodies- you can add a large amount of proteins to the complex.
