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Nucleic Acids Research, 1985, Vol. 13, No. 23 8293-8304
© 1985

Articles

Cloning of the human glucocorticoid receptor cDNA

Manjapra Variath Govindan, Martine Devic, Stephen Green, Hinrich Gronemeyer+ and P. Chambon

Laboratoire de Génétique Moléculaire des Eukaryotes du CNRS, Unité 184 de Biologie Moléculaire et de Génie Génétique de 1'INSERM, Faculté de Médecine 11 rue Humann, 67085 Strasbourg Cédex, France

Received October 24, 1985. Accepted November 12, 1985.

We show that the human glucocorticoid receptor (GR), isolated from the breast cancer cell line MCF-7, has an apparent molecular weight identical to that of rat liver GR (94 kDa) and reacts with antibodies raised against the latter. These antibodies were used to clone cDNA sequences corresponding to the human GR from a {lambda}gt11 expression library constructed using MCF-7 poly(A)+ RNA. Three non-homologous cONA clones with Inserts of 125, 220 and 350 bp, which express epitopes recognised by the rat liver GR antibodies, were isolated. Rat liver GR antibodies, immunopurified using the immobilised purified ß-galactos1dase fusion proteins, detect partially purified rat liver and human GRs on Western blots. In addition, these antibodies immuno-adsorb rat liver and human GRs affinity-labelled with [3H] triamcinolone acetonide. Northern blot analysis, using all three cDNA probes, reveals the presence of a major MCF-7 poly(A)+ RNA species of approximately 7 kb.

+Present address: Ludwig Institut für Krebsforschung, Inselspital 3010 Berne, Suisse


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