Soyika Richardson
Expressing Mammalian Signaling Proteins in Bacteria
Rhodopsin has been the only G-protein coupled receptor to ever have its X-ray crystal structure solved. Due to its availability in large quantities, inexpensiveness, and simple purification methods, the bovine rhodopsin protein has been frequently used in obtaining diffraction-quality 3-D crystals. In our experiments, the bovine rhodopsin protein has been cloned into the E. coli UT5600 plasmid using forward and reverse primers designed and purchased from Invitrogen. Plasmid DNA from these cells has been purified (using a Promega kit) in order to analyze the direction in which the protein has been inserted.