Affinity labelling and cloning of steroid and thyroid hormone receptors / edited by H. Gronemeyer.

Date:
[1988]
Catalogue details

Licence: Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)

Credit: Affinity labelling and cloning of steroid and thyroid hormone receptors / edited by H. Gronemeyer. Source: Wellcome Collection.

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    Ch. 16] Analysis of the human glucocorticoid receptor gene promoter 299 1985). The ability of the purified glucocorticoid receptor to interact with defined sequences in different hormonally regulated genes has been well documented (Payvar et al., 1981; Govindan et al., 1982; Geise et al., 1982; Pfahl, 1982; Scheidereit et al., 1983; Scheidereit and Beato, 1984; Karin et al., 1984; Renkawitz et al., 1984). Studies of steroid hormone receptors are of paramount interest not only for elucidating how the receptor interacts specifically with its Ugand but also for understanding the mechanism leading to the regulation of transcription through specific DNA-protein interactions. To investigate the regulation of the expression of the human glucocorticoid receptor gene at the molecular level, we have identified and characterized the 5' flanking region of this gene. II. DETERMINATION OF THE mRNA 5' END A. Southern analysis The genomic organization of the human glucocorticoid receptor gene was analyzed by digestion of human MCF-7 DNA with several restriction enzymes followed by Southern analysis (Fig. 1). The same blot was hybridized with nick-translated human 3 о cr-^— CLtOh-Q-OÛ Xiûcû >raw inc7>._ (ICO Q.COI— Q.ÛÛ X шаз Fig. 1 — Southern analysis of the human glucocorticoid receptor gene. Human genomic DNA (30 /xg) was digested with one of the enzymes PvuII, Sad, TaqI, PstI, Bglll, Hindlll, EcoRI or BamHI and electrophoresed on a 1.5% agarose gel. The DNA was transferred to diazobenzyl- oxymethyl (DBM) paper and hybridized with the nick-translated HGR1-1630 and HGR1-2850 cDNA probes. The position of the DNA size markers are indicated in kilobases.