Patent ID: 6015677
Filing Date: 2000-01-18
Classification: C07H,C07K,C12N,C12Q,G01N

Abstract:
An assay method for identifying a chemical agent which modifies the enzymatic activity of a mammalian cyclic GMP stimulated nucleotide phosphodiesterase said method comprising:(a) stably transforming with a polynucleotide encoding a mammalian cyclic GMP stimulated nucleotide phosphodiesterase selected from the group consisting of:(i) a polynucleotide encoding the amino acid sequence of SEQ ID NO: 39;(ii) a polynucleotide encoding the amino acid sequence of SEQ ID NO: 43;(iii) a polynucleotide encoding the amino acid sequence of SEQ ID NO: 45;(iv) a polynucleotide which hybridizes under stringent hybridization conditions to an antisense strand of the DNA insert in vector p3CGS-5 (A.T.C.C. 68579), pHcgs6n (A.T.C.C. 68962), pGSPDE6.1 (A.T.C.C. 68583), pGSPDE7.1 (A.T.C.C. 68585), or pBBCGSPDE-5 (A.T.C.C. 68578); and(v) a polynucleotide encoding the same polypeptide as (iv) by means of degenerate codons;a eukaryotic host cell having a heat-shock sensitivity phenotypic character susceptible to alteration, such that upon expression of said polynucleotide sequence said cell exhibits an altered heat-shock phenotype;(b) growing the transformed host cell formed in step (a) in a nutrient medium under conditions allowing expression of said polynucleotide sequence in said transformed host cell accompanied by the corresponding alteration in the transformed host cell phenotype;(c) contacting the transformed host cells grown according to step (b) with a chemical agent to be assayed; and(d) determining if the chemical agent step (c) modifies the enzymatic activity of the enzyme of step (a) by determining if the altered heat-shock phenotype of the cells of step (c) has been modified.