南移养殖仿刺参（StichopusjaponicusSelenka）铁蛋白基因的克隆及表达特征分析

采用cDNA文库、RACE、荧光定量PCR和Westernblot技术，克隆了南移养殖仿刺参铁蛋白基因的全长序列，并对其表达特征进行了研究。结果表明，南移养殖仿刺参铁蛋白cDNA全长1222bp，包括187bp的5’UTR；513bp的3’UTR和编码173个氨基酸残基的522bp的开放阅读框。诸如铁结合序列标签和铁调...

CLONING AND CHARACTERIZATION OF FERRITIN GENE FROM SOUTH CULTURED STICHOPUS JAPONICUS

With the approaches of cDNA library, RACE, fluorescent real-time quantitative PCR and western blot, the full-length ferritin cDNA was identified and characterized in South cultured Stichopus japonicus (denoted as SjFER) in the present study. All these results indicated: 1) The cDNA of SjFER was of 1222bp, consisting of a 5'UTR of 513bp with a putative iron regulatory element (IRE), a 3'UTR of 187bp, and a complete open reading frame of 522bp encoding a polypeptide with 173 amino acid residues. The conserved motifs for ferritin including iron binding signature, H-specific ferroxidase center and phosphorylation site were totally found in the deduced amino acid of SjFER. 2) SjFER was an important molecule in the species environmental adaption. The expression level of SjFER in South sea cucumber was two times higher than that from North in the tissue of muscle, while no difference occurred in tissue of respiratory tree. 3) The ployclonal antibodies generated from the recombinant product of SjFER could be specifically identified not only the recombinant product, but also the native protein from muscles, and should be used for next functional validation.