›› 2017, Vol. 29 ›› Issue (8): 1281-1289.DOI: 10.3969/j.issn.1004-1524.2017.08.08

• Animal Science • Previous Articles     Next Articles

Cloning and sequence analysis of growth hormone gene and its flanking region in topmouth culter (Culter alburnus Basilewsky)

LIU Shili1, 2, JIA Yongyi1, JIANG Wenping1, CHI Meili1, CHENG Shun1, ZHAO Jinliang2, GU Zhimin1, *, FU Jianjun3   

  1. 1. Agriculture Ministry Key Laboratory of Healthy Freshwater Aquaculture / Key Laboratory of Freshwater Aquatic Animal Genetic and Breeding of Zhejiang Province, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China;
    2. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China;
    3. Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China
  • Received:2017-03-17 Online:2017-08-20 Published:2017-09-06

Abstract: Using conserved primers and PCR, a gene encoding topmouth culter (Culter alburnus Basilewsky) growth hormone (CaGH) was amplified and sequenced. The gene spans 5 966 bp, including 2 282 bp of 5'- and 2 036 bp of 3'-flanking sequences and a 1.7-kb transcription unit comprised of five exons and five introns. (AAT)8 and (TTC)5T(TAA)8 microsatellite sequences were found in the 5'- and 3'-flanking regions, respectively. The upstream region contains TATA boxes, and binding sites of important transcription factors such as Pit-1, Pit-1a, CREB, AP1, GR, HNF-3, and HNF-3B. The five exons in Culter alburnus were 64 bp, 140 bp, 117 bp, 162 bp and 255 bp in length, respectively. The complete coding sequence was 603 bp and encodes a protein with a 22 amino acid signal peptide and a 178 amino acid mature peptide. Five conserved Cys residues (Cys71, Cys135, Cys173, Cys190, and Cys198) and two possible sites of N-glycosylation (residues 145 and 197) were detected in the GH polypeptide. The amino acid sequence of GH in Culter alburnus was identical to that in Megalobrama amblycephala, and only one amino acid residue differs from Ctenpharyngodon idellus. The phylogenetic relationships among the GH amino acid sequences in fish were in accord with traditional classification. The lengths of the four introns in Culter alburnus GH gene were 229 bp, 103 bp, 565 bp and 103 bp, respectively. The variation of the introns among species was greater than that of the exons, and the variation of the third intron is the highest. These results provided the molecular basis for study of function and transcriptional regulation of the GH gene in C.alburnus, as well as the temporal expression in different developmental stages and at various nutritional levels.

Key words: Culter alburnus, growth hormone gene, clone, sequence analysis

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