American Cyanamid Company

Laura A Price

Mark Henry Pausch

Donald R Kirsch

Bradley Alton Ozenberger

John Richard Hadcock

Deborah Tardy Chaleff

Eileen Marie Kajkowski

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.

Recombinant expression vectors for expression of heterologous proteins

A first aspect of the present invention is directed to a transformed yeast cell containing a first heterologous DNA sequence which codes for a G protein-coupled receptor, for example, the somatostatin receptor, and a second heterologous DNA sequence which codes for a G protein .alpha. subunit or portions thereof fused to DNA sequences from the yeast G protein a subunit. A second aspect of the present invention is a transformed yeast cell containing a heterologous DNA sequence which codes for a G protein coupled receptor. A third aspect of the present invention is a method of assaying compounds to determine effects on cell growth.

Transformed yeast cells expressing heterologous G-protein coupled

A first aspect of the present invention is the isolation and characterization of invertebrate (i.e. insect and nematode) potassium channel genes. The present invention provides for the isolation of complementary DNA fragments from Drosophila melanogaster and Caenorhabditis elegans which encode conserved amino acid sequence elements unique to the potassium channel gene family. A yeast expression technology is employed to clone cDNAs from Drosophila melanogaster and a hybridization approach is utilized to isolate additional cDNAs from Caenorhabditis elegans. Using the yeast expression technology, a single 2463 base pair cDNA fragment designated Dm ORF1 is isolated.

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Langhorne, PA, United States of America

Laura A Price