Expression of G protein coupled receptors in yeast

Details Expression of G protein coupled receptors in yeast Expression of G protein coupled receptors in yeast

2004-11-23

2008-08-19

Ulm, John (Department: 1649)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

C435S007310, C435S069100, C435S252300

Reexamination Certificate

active

07413876

ABSTRACT:
Disclosed is a transformed yeast cell containing a first heterologous DNA sequence which codes for a mammalian G protein-coupled receptor and a second heterologous DNA sequence which codes for a mammalian G protein α subunit (mammalian Gα). The first and second heterologous DNA sequences are capable of expression in the cell, but the cell is incapable of expressing an endogenous G protein α-subunit (yeast Gα). The cells are useful for screening compounds which affect the rate of dissociation of Gαfrom Gβγin a cell. Also disclosed is a novel DNA expression vector useful for making cells as described above. The vector contains a first segment comprising at least a fragment of the extreme amino-terminal coding sequence of a yeast G protein-coupled receptor. A second segment is positioned downstream from the first segment (and in correct reading frame therewith), with the second segment comprising a DNA sequence encoding a heterologous G protein-coupled receptor.

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