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| Cloning,Characterization and Expression of Full-length cDNA of a Copper/Zinc Superoxide Dismutase Gene in Sea Cucumber (Apostichopus j aponicus) |
| GAO Shan,DONG Ying,WANG Bai,YANG Ai-fu,CHEN Zhong,GUAN Xiao-yan,JIANG Jing-wei,JIANG Bei,SUN Hong-juan,ZHOU Zun-chun |
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Abstract In this study ,the full‐length cDNA of Cu/Zn‐SOD gene as an immune‐related gene was cloned and characterized in sea cucumber (A postichopus japonicus) .It was found that the Cu/Zn‐SOD gene had full‐length of 1 500 bp including a 129 bp 5’‐untranslated region (UTR) ,459 bp ORF encoding 152 amino acids ,and a 912 bp 3’‐UTR with predicted molecular weight of 17 .47 ku .Four copper binding sites (His‐44 ,‐46 ,‐61 ,and‐118) and four Zn binding sites (His‐61 ,‐69 ,‐78 ,and Asp‐81) ,a conserved disulfide bond (Cys‐55 ,‐144 ) , and two conserved Cu/Zn‐SOD signatures were identified in the deduced amino acid sequence in the Cu/Zn‐SOD .The deduced amino acid sequence showed 71% amino acid sequence identity to the Cu/Zn‐SOD in Leptopilinaboulardi and Bemisia tabaci .Phylogenetic analysis revealed that the Cu/Zn‐SOD gene in the sea cucumber was close to that in purple sea urchin Strongylocentrotus purpuratus in invertebrate cluster .Quantitative real‐time PCR analysis showed that the Cu/Zn‐SOD gene was expressed in all the tested tissues including body wall ,feet ,coelomocytes ,respiratory tree ,muscle ,and intestine , the maximal expression level in feet . The expression of the Cu/Zn‐SOD mRNA was down‐regulated significantly in the coelomocytes 4—12 h after bacterial lipopolysaccharide (LPS) challenge ,indicating that the Cu/Zn‐SOD gene would be one of the immune‐related factor .
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Received: 25 May 2014
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