Patent ID: 6432652
Filing Date: 2002-08-13
Classification: C07K

Abstract:
A method to screen a candidate substance for opioid agonist activity, which method comprises:(a) incubating cells displaying opioid receptor in the presence and absence of the candidate substance under conditions suitable for detection of activation of said opioid receptor, and (b) detecting the amount of activation of said opioid receptor in the presence as compared to the absence of said candidate substance; whereby an increase in the amount of activity in the presence, as compared to the absence of said candidate substance indicates that the substance is an agonist for said opioid receptor wherein said opioid receptor is produced and displayed on said cells by a method which comprises culturing said host cells which have been modified to contain an expression cassette which produces said opioid receptor in the host cells, said expression cassette comprising a nucleotide sequence encoding said opioid receptor operably linked to control sequences operable to provide expression in said host cells, and wherein said opioid receptor is a mammalian opioid receptor which is able to transduce a signal when contacted with an opioid agonist and is encoded by a nucleotide sequence which hybridizes under conditions of high stringency to the nucleotide sequence of SEQ. ID. NO: 7 or the complement thereof, wherein said conditions of high stringency comprise final wash steps of washing twice in 40 mM NaPO4, pH 7.2; 0.5% BSA; 5% SDS; and 1 mM EDTA for one hour at 68Â° C. followed by final wash steps of washing twice in 40 mM NaPO4, pH 7.2; 1% SDS; and 1 mM EDTA for one hour at 68Â° C.