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

C - Chemistry, Metallurgy

12

N

C12N 15/12 (2006.01) C07K 14/39 (2006.01) C07K 14/47 (2006.01) C07K 14/705 (2006.01) C12N 1/19 (2006.01) C12N 15/62 (2006.01) C12N 15/81 (2006.01) C12Q 1/34 (2006.01) C12Q 1/68 (2006.01) G01N 33/566 (2006.01)

Patent

CA 2092717

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 a subunit (mammalian G.alpha.). 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 .alpha.-subunit (yeast G.alpha.). The cells are useful for screening compounds which affect the rate of dissociation of G.alpha. from G.beta..gamma. in 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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