Vectors for expression of G protein coupled receptors in yeast
AssigneeDuke University
InventorsKlim King, Henrik G. Dohlman, Marc G. Caron, Robert J. Lefkowitz
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AssigneeDuke University
InventorsKlim King, Henrik G. Dohlman, Marc G. Caron, Robert J. Lefkowitz
Estimating the term…
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 .alpha. subunit (mammalian G.sub..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.sub..alpha.). The cells are useful for screening compounds which affect the rate of dissociation of G.sub..alpha. from G.sub..beta..tau. 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.
A DNA expression vector capable of expressing a transmembrane protein into the cell membrane of a yeast cell, said yeast cell lacking an endogenous G protein .alpha. subunit and expressing a mammalian Gs .alpha. subunit, said vector comprising:
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