HOST CELLS EXPRESSING MUTANTS OF A NATURAL G-PROTEIN COUPLED RECEPTOR (GPCR); SACCHAROMYCES CEREVISIAE STE2 GENE, AND THEIR USE AS BIOSENSORS

摘要

A method for the preparation of cells (useful as biosensors) which present on their surface a mutant of a natural G-protein coupled receptor (GPCR), said mutant being capable of binding a desired ligand which the natural receptor cannot, which method comprises: generating a diverse collection of replicable expression plasmids which comprise (i) a coding sequence coding for a mutated form of a natural GPCR, wherein the mutation is of one or more amino acids in the transmembrane and/or extracellular domains of the natural GPCR, and (ii) expression control sequences operatively linked to said coding sequence to control expression of the mutant GPCR; transforming cells of a host organism with said diverse collection of plasmids; culturing the resultant collection of transformed cells for a sufficient time to permit expression of mutant GPCR from plasmids replicating within such cells; contacting the cultured cells with the desired ligand; and isolating cells wherein the surface presented mutant GPCR expressed by such cells binds to the desired ligand.