277 zyxwvutsr Gene, 143 (1994)277-280 0 1994 Elsevier Science B.V. All rights reserved. 0378-l 119/94/$07.00 GENE 07894 A dog growth hormone cDNA codes for a mature protein identical to pig growth hormone (Canisfamiliaris; pituitary gland; polymerase chain reaction; nucleotide sequence; bacterial expression) Jorge A. Ascacio-Martinez and Hugo A. Barrera-Saldafia Departamento de Bioquimica, Facultad de Medicina, Universidad Autdnoma de Nuevo Ledn., Monterrey, N.L., 64000, MJxico Received by F. Bolivar: 10 August 1993; Revised/Accepted: 17 December/20 December 1993; Received at publishers: 14 February 1994 SUMMARY Although methods for purification of dog (Canisfamiliaris) growth hormone (cfGH) were described in the late Sixties, the cloning of its cDNA has not been achieved until now. In order to clone the cfCH cDNA, we capitalized on the high degree of nucleotide sequence conservation among mammalian GH genes to design a pair of consensus oligodeoxyri- bonucleotide primers. With these, and starting with dog pituitary gland total RNA, we specifically amplified the cfGH cDNA using the reverse transcription-polymerase chain reaction. Its coding sequence (651 bp), as well as its 3’ un- translated region (101 bp), resemble those of a typical mammalian GH cDNA. Interestingly, its encoded mature protein is identical to pig growth hormone (pGH). INTRODUCTION Mammalian growth hormones (GHs) are single 22-kDa polypeptides typically of 190 aa that present structural and functional similiarities (Catt et al., 1967; Sherwood, 1967; Niall et al., 1971; 1973). Complete nt sequences have been determined for GH genes or their cDNAs from the following mammalian species: man (DeNoto et al., 1981; Seeburg, 1982), rat (Barta et al., 1981; Page et al., 1981), cattle (Woychik et al., 1982; Seeburg et al., 1983), pig Correspondence to: Dr. H.A. Barrera-Saldafia, Departamento de Bioquimica, Facultad de Medicina de la Universidad Autonoma de Nuevo Leon, A.P. 3&-4125, Monterrey, N.L., 64000, Mexico. Tel. and Fax (52-8) 333-7747; e-mail:habarrer@academ08.mty.itesm.mx Abbreviations: aa, amino acid(s); bp, base pair(s); BGal, S-galactosidase; bGH, bovine GH; cDNA, DNA complementary to mRNA; cfGH, Canis familiaris GH; dGTP, deoxyguanosine triphosphate; dITP, deoxiino- sine triphosphate; E., Escherichia; GH, growth hormone; GH, gene or cDNA encoding GH; hGH, human GH; IPTG, isopropyl-B-D- thiogalactopyranoside; kb, kilobase or 1000 bp; MBP, maltose- binding protein; nt, nucleotide(s); oligo, oligodeoxyribonucleotide; PCR, polymerase chain reaction; pGH, pig GH; re-cfGH, recombinant cfGH; SDS, sodium dodecyl sulfate. SSDI 0378-1119(94)00105-2 (Seeburg et al., 1983), sheep (Warwick et al., 1989), goat (Yamano et al., 1988) and mink (Shoji et al., 1990). Deletions of the GH gene causes autosomal recessive dwarfism in man (McKusick, 1978), mice (Nicholas, 1978; Either and Beamer, 1976), German Shepherd and in other breeds of dogs (Nicholas, 1978; Andresen et al., 1974; Andresen and Willeberg, 1976a,b). Therapy for pituitary dwarfism relies on the admin- istration of GH (Feldman and Nelson, 1987). Within 4 to 6 weeks after initiating therapy, adult dogs show re- growth of hair and thickening of the skin. Unfortunately, dog (Canisfamiliaris) GH (cfGH) preparations are diffi- cult to obtain and therefore expensive. Perfect immuno- logical cross-reactivity with pig GH (pGH) has been reported for cfGH (Cocola et al., 1976) and indeed bovine GH (bGH) and pGH have been used experimentally to treat canine pituitary dwarfism (Feldman and Nelson, 1987; Eigenmann, 1981). Phenylalanine comprises the C- and N-terminal residues of cfGH, as is the case for porcine, bovine, ovine and human hormones. The aa composition and other chemical and immunological properties of cfGH were found to be very similar to those of pGH (Scott et al., 1978).