Chemistry: molecular biology and microbiology – Vector – per se
Patent
1995-06-06
1998-12-08
Walsh, Stephen
Chemistry: molecular biology and microbiology
Vector, per se
435 691, 530350, C12N 1563, C12N 1512, C07K 14705
Patent
active
058468196
ABSTRACT:
The present invention is directed to a expression vectors and yeast cells transformed therewith containing a first heterologous nucleotide sequence which codes for a G protein-coupled receptor, for example, the somatostatin receptor, and a second nucleotide sequence which codes for all or a portion of a G protein .alpha..beta..gamma. complex. Said heterologous protein is physically expressed in a host cell membrane in proper orientation for both stereoselective binding of ligands, as well as functional interaction with G proteins on the cytoplasmic side of the cell membrane. In some embodiments, a nucleotide sequence encoding a heterologous or chimeric G.alpha. protein is expressed in conjunction with nucleotide sequences from the yeast G protein .beta..gamma. subunits. A second aspect of the present invention provides expression vectors and transformed yeast cells encoding chimeric yeast/heterologous G protein coupled receptors. A third aspect of the present invention is directed to methods of assaying compounds using these expression constructs and yeast cell expression systems to determine the effects of ligand binding to the heterologous receptors expressed in the systems.
REFERENCES:
patent: 4925791 (1990-05-01), Nasmyth et al
patent: 5284746 (1994-02-01), Sledziewski et al.
patent: 5482835 (1996-01-01), King et al.
H.G. Dohlman, J. Thorner, M.G. Caron, R.J. Lefkowitz--Model Systems For The Study of Seven-Transmembrane-Segment Receptors, Annu. Rev. Biochem. 1991. 60:653-88.
W.C. Probst, L.A. Snyder, D.I. Schuster, J. Brosius, S.C. Sealfon--Sequence Alignment of the G Protein-Coupled Receptor Superfamily, DNA and Cell Biology, vol. 11, No. 1, 1992, pp. 1-20.
Klim King, Henrik G. Dohlman, Jeremy Thorner, Marc G. Caron, Robert J. Lefkowitz--Control of Yeast Mating Signal Transduction by a Mammalian .beta..sub.2 -Adrenergic Receptor and G.sub.s .alpha. Subunit, Science, vol. 250, pp. 121-123.
Ira Herskowitz and Lorraine Marsh, Conservation of a Receptor/Signal Transduction System, Cell, vol. 50, 995-996, Sep. 25, 1987.
Stafano Marullo, Colette Delavier-Klutchko, Jean-Gerard Guillet, Alain Charbit, Arthur Donny Strosberg and Laurent Jean Emorine, Expression of Human .beta..sub.1 and .beta..sub.2 Adrenergic Receptors in E. Coli as a New Tool for Ligand Screening, Bio/Technology, vol. 7, Sep. 1989, pp. 923-927.
Gretchen McCaffrey, Fiona J. Clay, Kimberly Kelsay and George F. Sprague, Jr., Identification and Regulation of a Gene Required for Cell Fusion during Mating of the Yeast Saccharomyces cerevisiae, Molecular and Cellular Biology, vol. 7, No. 8, 1987, pp. 2680-2690.
Joshua Trueheart, Jef D. Boeke, and Gerald R. Fink, Two Genes Required for Cell Fusion during Yeast Conjugation: Evidence for a Pheromone-Induced Surface Protein, Molecular and Cellular Biology, Jul. 1987, pp. 2316-2328.
J.W. Kronstad, J.A. Holly, and V.L. MacKay, A Yeast Operator Overlaps an Upstream Activation Site, Cell, vol. 50, 369-377.
Chaleff Deborah T.
Hadcock John R.
Kajkowski Eileen M.
Kirsch Donald R.
Ozenberger Bradley A.
American Cyanamid Company
Basham Daryl A.
Matthews Gale F.
Walsh Stephen
LandOfFree
Recombinant expression vectors for expression of heterologous pr does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Recombinant expression vectors for expression of heterologous pr, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Recombinant expression vectors for expression of heterologous pr will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-176562