Yeast receptor and g-protein fusion protein

Details Yeast receptor and g-protein fusion protein Yeast receptor and g-protein fusion protein

C - Chemistry, Metallurgy

12

N

C12N 15/62 (2006.01) C07K 14/395 (2006.01) C07K 14/705 (2006.01) C12N 1/19 (2006.01) C12N 15/31 (2006.01) C12N 15/52 (2006.01) C12N 15/81 (2006.01) C12Q 1/68 (2006.01) G01N 33/566 (2006.01)

Patent

CA 2232733

The invention provides protein fusions between the C-terminus of heterotrimeric G-protein-coupled receptors and the N-terminus of either wild type or mutant G-alpha proteins of the yeast Saccharomyces cerevisiae. Methods are described for creating DNA constructs that encode such fusion protein, assays for correct expression of such fusion molecules in yeast, and assays for the coupling of such fusion molecules to the pheromone-induced signal transduction pathway of yeast. Furthermore, the invention encompasses yeasts expressing the fusion proteins and methods for screening compounds for activity as agonists or antagonists of seven-transmembrane receptor function.

L'invention se rapporte à des fusions de protéines entre l'extrémité C-terminale de récepteurs hétérotrimériques couplés à la protéine G et l'extrémité N-terminale de protéines G-alpha du type sauvage ou mutant de la levure Saccharomyces cerevisiae. L'invention concerne des procédés pour créer des ADN de synthèse qui codent une telle protéine de fusion; des dosages pour exprimer correctement de telles molécules de fusion dans la levure; et des dosages pour coupler de telles molécules de fusion au processus de transduction d'un signal induit par phéromone d'une levure. En outre, l'invention concerne les levures exprimant les protéines de fusion et les procédés permettant de cribler des composés agissant comme agonistes ou antagonistes d'un récepteur à sept transmembranes.

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