Isolation, cDNA cloning, and growth promoting activity of rabbitfish (Siganus guttatus) growth hormone

We report the isolation, cDNA cloning, and growth promoting activity of rab bitfish (Siganus guttatus; Teleostei; Perciformes; Siganidae) growth hormon e (GH). Rabbitfish GH was extracted from pituitary glands under alkaline co nditions, fractionated by gel filtration chromatography on Sephadex G-100, and purified by highperformance liquid chromatography. The fractions contai ning GH were identified by immunoblotting with bonito GH antiserum. Under n onreducing conditions, the molecular weight of rabbitfish GH is about 19 kD a as estimated by SDS-PAGE. The purified hormone was potent in promoting gr owth in rabbitfish fry. Weekly intraperitoneal injections of the hormone si gnificantly accelerated growth. This was evident 3 weeks after the start of the treatment, and its effect was still significant 2 weeks after the trea tment was terminated. Rabbitfish GH cDNA was cloned to determine its nucleo tide sequence. Excluding the poly (A) tail, rabbitfish GH cDNA is 860 base pairs (bp) long. It contained untranslated regions of 94 and 175 bp in the 5' and 3' ends, respectively. It has an open reading frame of 588 bp coding for a signal peptide of 18 amino acids and a mature protein of 178 amino a cid residues. Rabbitfish GH has 4 cysteine residues. On the amino acid leve l, rabbitfish GH shows high identity (71-74%) with GHs of other perciforms, such as tuna, sea bass, yellow tail, bonito, and tilapia, and less (47-49% ) identity with salmonid and carp GHs. (C) 2000 Academic Press.