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Molecular and expression characterization of growth hormone/prolactin family genes in the Prenant’s schizothoracin

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Abstract

The growth hormone/prolactin family genes are major participants in control of several complex physiologic processes, including growth, reproduction, and metabolism. In this study, total RNA was isolated from the pituitary of the Prenant’s schizothoracin (Schizothorax prenanti), and SMART cDNA was synthesized from 100 ng total RNA. The full-length cDNAs of three GH/PRL family genes have been cloned and sequenced from the SMART cDNA library. The growth hormone of S. prenanti (SpGH) cDNA contains 1238 base pairs (bp) and encodes 215 amino acid (aa) residues; the prolactin (SpPRL) cDNA contains 1124 bp and encodes 209 aa residues; the cDNA sequence of somatolactin (SpSL) is 1290 bp in length, and encodes for a peptide of 234 aa residues. Although there is only about 22% aa sequence identity between the three deduced proteins, SpGH, SpPRL and SpSL, overall, the C-terminal region shows a higher identity. Multiple aa sequence alignments indicated the deduced SpGH, SpPRL and SpSL show high identities to that of goldfish and zebrafish, respectively. RT-PCR analysis revealed that the three hormones displayed different tissue distribution patterns. SpPRL transcripts were detected only in the pituitary of the Prenant’s schizothoracin. SpSL was more widely distributed than SpPRL, high mRNA levels of SpSL were detected in the pituitary and low mRNA levels of SpSL were found in liver, kidney and heart. SpGH was expressed in nearly all tissues detected with the highest expression level in the pituitary.

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Acknowledgments

We thank Dr. Taiming Yan (Sicuan Agriculture University) for sampling. This work was financially supported by the director fund of Yangtze River Fisheries Research Institute, CAFS, and National Nonprofit Institute Research Grant of Freshwater Fisheries Research Center, Chinese Academy of Fisheries (CAFS), and the foundation of Key Laboratory of Freshwater Biodiversity Conservation and Utilization, Ministry of Agriculture of China (LFBCU0704).

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Correspondence to Xihua Chen.

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11033_2010_592_MOESM1_ESM.doc

Fig. 1 Nucleotide and the deduced amino acids sequence of the SpGH a, and SpPRL b, and SpSL c. The nucleotide and amino acid residue number are shown on the left. Amino acids of the signal peptide are underlined and given negative numbers and the first amino acid of each mature protein is bolded and numbered as +1. Consensus polyadenylation signals are boxed. Putative N-linked glycosylation sites at Asn residue are highlight and Cys residues in mature peptide are shaded. The primers used in each RT-PCR analysis were marked by forward and reverse arrows under the nucleotide sequence. The nucleotide sequences of SpGH, and SpPRL, and SpSL have been deposited in the GenBank nucleotide database, under accession no. GQ845447, GQ845448, and GQ845449, respectively. Fig. 2 Alignment of the amino acid sequences of the SpGH a, SpPRL b, and SpSL c and other vertebrates. Multiple alignments were performed with the MAP method at BCM Search Launcher web servers and the printing output was shaded by BOXSHADE 3.21. Identical residues are in black, conservative substitutions are in gray. The Cys residues are shaded and numbered accordingly. Numbers of residues are shown at the left margin. The two black arrows indicate positions for degenerate primers design. The aa sequences used in this study were extracted from GenBank databases as listed in Fig. 2. Aa Anguilla anguilla, Xl Xenopus laevis, Gg G gallus gallus, Hs Homo sapiens, Dr Danio rerio, Ok Oncorhynchus keta, Sa Sparus aurata, Ca Carassius auratus, Pg Prionace glauca, Ag Acipenser gueldenstaedtii, As Acipenser sinensis, Po Paralichthys olivaceus, Pa Protopterus annectens, On Oreochromis niloticus, Ol Oryzias latipes, Sp Schizothorax prenanti. (DOC 1832 kb)

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Li, C., Chen, X., Zhang, Y. et al. Molecular and expression characterization of growth hormone/prolactin family genes in the Prenant’s schizothoracin. Mol Biol Rep 38, 4595–4602 (2011). https://doi.org/10.1007/s11033-010-0592-2

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