Method for identifying and/or isolating genes which code for membrane receptors
AssigneeEnichem Partecipazioni S.p.A.
InventorsGabriella Galli, Rafaele Matteoni
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Abstract
A method for the analysis and isolation of genes which express membrane receptors was developed. Cells of E. coli transformed with proper vectors, from phage, as well as from plasmids, containing the cDNA coding for human .beta.-adrenergic receptors, are grown on slabs of LB/agar; the bacterial colonies, once transferred onto nitrocellulose filters, show specific activity for (.sup.125 I)-iodocyanopindolol (ICYP). The .beta.-adrenergic receptors expressed in bacteria retain their pharmacologic characteristics after being transferred on filters.
Claims4
A method for identifying and/or isolating DNA encoding G-coupled membrane receptors, comprising:
- preparing a vector encoding a membrane receptor which naturally occurs coupled to a G-protein;
- transforming E. coli cells using said vector;
- adsorbing bacterial colonies derived from said transformed bacterial cells on a filter of nitrocellulose;
- incubating said colonies on said filter with a labeled ligand;
- identifying those colonies containing said DNA by their ability to bind said ligand;
- and, optionally, isolating said DNA.
Dependencies are inferred from the claim text.