Production of olfactory receptors using commercial E. coli cell-free systems

Abstract

The first bottleneck in olfactory receptor (OR) structural and functional studies is to produce sufficient quantities of soluble, functional, and stable receptors. Other production systems have been used and summarized in other chapters of this book. We here show that commercial cell-free in vitro translation systems can be used to produce milligrams of soluble and functional olfactory receptors within several hours directly from plasmid DNA with select optimal detergents. These olfactory receptors can be purified using immunoaffinity 1D4 monoclonal antibody rhodopsin-tag and gel filtration, and can be analyzed using gel electrophoresis and with other standard techniques. The olfactory receptors and other scent-related receptors produced by the cell-free method fold properly and are able to bind their odorants.