Gene, 163 (1995) 331-332 © 1995 Elsevier Science B.V. All rights reserved. 0378-1119/95/$09.50 GENE 09075 Letter to the Editor Sequence of the :5' regulatory domain of the gene encoding the rat 132-adrenergic receptor (5' Flanking region; nucleotide sequence; glucocorticoid; thyroid hormone; restriction map) 331 Liu Jiang and George Kunos Department of Pharmacology ana Toxicology, Medical College of Virginia, Virginia Commonwealth University, Richmond, VA 23298, USA Received by J.A. Engler: 17 March 1995; Revised/Accepted: 21 April/26 April 1995; Received at publishers: 18 May 1995 SUMMARY Restriction mapping and sequence analysis of the gene encoding the rat ~2 adrenergic receptor (132AR) revealed an error affecting most of the Y-flanking domain, located between - 192 and - 2245 bp in the only published sequence of this gene [Buckland et al., Nucleic Acids Res. 18 (1990) 682]. The correct sequence lacks a TATA box, contains 67% G + C residues in the first 500 nucleotides upstream from the start codon, and contains putative glucocorticoid, thyroid hormone and cAMP-response elements. The 132-adrenergic receptor (~2AR) is a member of the family of G-protein-coupled receptors, and it plays an important role in mediating the sympathoadrenal response to stress. The only published sequence of the rat fl2AR gene (Buckland et al., 1990) differs in the 5' regulatory domain from that in other species. Therefore, as the first step toward characterizing the structural ele- ments involved in its transcription, we have isolated the rat fl2AR gene. A rat liver genomic library was screened with a 2.1-kb rat fl2AR cl-)NA probe containing the entire coding region (Gocayne et al., 1987). A positive clone identified under stringent hybridizing conditions was sub- cloned into pGEM(-)77 plasmids. Restriction mapping by restriction enzyme digestion and Southern blotting have established that the 5.5-kb fragment contains the entire coding region plu~; approximately 3.3 kb of the 5' M H A P BIB2E B A D A C Kp S Ap X P A c S/Ap PH P X I II III I Correspondence to: Dr. G. Kunos, Department of Pharmacology and Toxicology, Virginia Commonwealth University, Box 980613, Richmond, VA 23298, USA. Tel. (1-804) 828-2073; Fax (1-804) 828-2117; e-mail: GKunos@gems.vcu.edu Abbreviations: bp, base pair(s); 132AR, 132-adrenergic receptor; fl2AR, gene encoding [~2AR; CRE, cAMP response element(s); GRE, glucocor- ticoid response element(s); kb, kilobase(s) or 1000 bp; nt, nucleotide(s); TRE, thyroid hormone respo~se element(s). I lkbl Fig. 1. Restriction mapping of the 5.5-kb clone containing the rat ~:AR- encoding gene. The shaded segment in the horizontal bar represents the coding region. The digestion products were separated on a 1.5% agarose gel and visualized by ethidium-bromide staining (top). Lanes: M, 1-kb DNA ladder; H, HindIII; A, AvaI; P, PstI; B1, BgII, B2 BglII; E, EcoRI; BA, BamHI; D, DraI; Ac, AccI; Kp, KpnI; S, Sinai; Ap, ApaI; X, XhoI. SSD1 0378-1119(95)00357-6