双棘黄姑鱼IGF2 基因克隆及其在卵巢发育中的作用研究

    IGF2 gene cloning and its function during the development of ovarian cycle in Protonibea diacantus

    • 摘要: 运用 RT-PCR 和 RACE 技术,首次克隆了双棘黄姑鱼(Protonibea diacanthus)的 IGF2 cDNA。双棘黄姑鱼 IGF2 cDNA 全长 842 bp,其中开放阅读框为 648 bp,5忆非翻译区为 125 bp,3忆非翻译区为 69 bp。 IGF2 前体mRNA 编码 215 个氨基酸,成熟肽为 71 个氨基酸残基的多肽。多重氨基酸比对结果显示,双棘黄姑鱼与其他鱼类 IGF2 氨基酸序列高度保守。 同源性分析表明,双棘黄姑鱼 IGF2 氨基酸序列与同属鲈形目的斜带石斑鱼、金头鲷、罗非鱼同源性分别为 95.8%、95.3%和 91.2%。 系统进化树结果与同源性分析结果相似,双棘黄姑鱼 IGF2基因首先与同属鲈形目的斜带石斑鱼、金头鲷、罗非鱼类聚,再与虹鳟、牙鲆、黄颡鱼的 IGF2 和斑马鱼 IGF2b基因聚为一支,最后与斑马鱼 IGF2a 类聚为鱼类 IGF2 基因。 组织分布结果表明,IGF2 基因在双棘黄姑鱼各组织中表达广泛。 RT-PCR 结果显示, 双棘黄姑鱼卵巢成熟期和退化期的 IGF2 表达量显著高于发育期, 推测IGF2 的表达可能促进了双棘黄姑鱼卵巢的成熟。

       

      Abstract: IGF cDNA of Protonibea diacanthus was obtained by using the method of RACE and RT-PCR for the first time in this study. The full length sequence is 842 bp, which contains 648 bp open reading frame (ORF), 125 bp 5untranslated region (UTR), and 69 bp 3' untranslated region. IGF2 precursor mRNA encodes 215 amino acids, while the mature polypeptides contains 71 amino acid residues. IGF2 of Protonibea diacanthus has a high homology with that of other fishes. It has 95.8%, 95.3%, 91.2% homology with Epinephelus coioides, Sparus aurata, Oreochromis niloticus, respectively. Phylogenetic tree analysis shows that the IGF2 of Protonibea diacanthus tends to cluster with E. coioides, S. aurata, O. niloticus of the same genus firstly, then clusteres with IGF2 of Oncorhynchus mykiss, Ietalurus Punetaus, Pelteobagrus fulvidraco and IGF2b of Danio rerio in turn. Finally IGF2a of Danio rerio is clustered in the same branch with bony fishesIGF2. Tissue distribution shows that IGF2 is widely expressed in multiple tissues of P. diacanthus. RT-PCR reveales that the highest expression of IGF2 of ovary is detected in maturation phase and regression phase than that development phase. IGF2 expression may relate with ovarian maturation of P. diacanthus.

       

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