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Issue:ISSN 1000-7083
          CN 51-1193/Q
Director:Sichuan Association for Science and Technology
Sponsored by:Sichuan Society of Zoologists; Chengdu Giant Panda Breeding Research Foundation; Sichuan Association of Wildlife Conservation; Sichuan University
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Your Position :Home->Past Journals Catalog->2014 Vol.33 No.6

Molecular Cloning, Sequence Analysis and Tissue Expression of Selenoprotein W in Common Carp
Author of the article:CHEN Wenbo1*, DONG Haiyan2, ZHANG Zhen1, JIANG Xiaoxue1
Author's Workplace:(1. Department of Biology, Institute of Resources and Environment, Henan Polytechnic University, Jiaozuo, Henan Province 454000, China; 2. Medicine Department, Huzhou Teachers College, Huzhou, Zhejiang Province 313000, China)
Key Words:Cyprinus carpio; selenoprotein W; cloning; tissue expression
Abstract:Selenoprotein was a kind of protein that contained selenocysteine which played an important role in animals. Although selenium (Se) was very important for fish’s growth, development, antioxidant and immunity, there were few studies about selenoproteins in fish, especially on the structure and function of selenoprotein (SelW). The full-length of SelW cDNA (ccSelW) was studied in common carp (Cyprinus carpio) by cloning from hepatopancreas through RT-PCR and RACE techniques. The results indicated that the ccSelW cDNA sequence was 704 bp, containing a 5′-untranslated region (UTR) of 47 bp, a 3′-UTR of 396 bp with a poly(A) tail, and an open reading frame of 261 bp encoding a polypeptide of 86 amino acids. And the ccSelW had several conserved features including the C-X-X-U motif and selenocysteine (Sec) in site 13. The tertiary structure model of ccSelW was constructed by SWISS-MODEL based on the template of mouse SelW. A β-α-β-β-β-α secondary structure pattern, which was similar to mouse, was also displayed. An 89 bp SECIS element, which formed a classic AUGA-AA-GA secondary structure, was identified in the 3′-UTR of ccSelW cDNA by using the program SECISearch 2.19. Because of the additional stem-loop structure on the top of the apical loop, the SECIS of common carp should be classified as type Ⅱ. Common carp had the highest homology to Danio rerio and Xenopus tropicalis (84.7%), but relatively lower to other species (54%~60%). The phylogenetic tree which was constructed by neighbor-joining method with 1000 replicates indicated that, ccSelW was clustered into the same branch with D. rerio and X. tropicalis. The result of semi-quantitative RT-PCR revealed that ccSelW mRNA was expressed in all detected tissues. The highest expression level was observed in testis, followed by hypothalamus and intestine. Our research enriched the knowledge of SelW in fish, and provided a foundation for illuminating its structure and physiological function.
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