In Vitro Analysis of PDZ-dependent CFTR Macromolecular Signaling Complexes

The overall goal of this procedure is to assemble A PDZ dependent cystic fibrosis transmembrane conductance regulator, or CFTR macromolecular signaling complex. In vitro to do this, the recombinant tagged fusion proteins are expressed in bacteria and purified. Next cell lysates are obtained from cultures that express CFTR or CFTR binding proteins.

Then the CFTR and binding proteins are combined in vitro to assemble the CFTR containing macromolecular signaling complex. Western blotting is then performed, which demonstrates a formation of CFTR macromolecular complexes and is then assessed via western blotting. The main advantage of this technique over existing methods is that we can directly detect and visualize the formation of a C CT R containing high-protein macromolecular complex individual.