黄喉拟水龟转铁蛋白基因的克隆以及表达特征分析

CLONING AND EXPRESSION PATTERN ANALYSIS OF TRANSFERRIN GENE IN YELLOW POND TURTLE

  • 摘要: 转铁蛋白在铁的新陈代谢中起着重要的作用,参与细菌感染后的免疫反应。研究克隆了黄喉拟水龟Tf基因组DNA全长序列,并对Tf基因的序列特征进行了分析。序列分析表明,黄喉拟水龟Tf是由两个相似结构域构成的单一肽链,每个结构域包含两个亚基,它们相互作用形成一个深的、亲水的铁结合位点。其基因组DNA由17个外显子和16个内含子组成,与其他脊椎动物Tf基因结构相似,显示了黄喉拟水龟Tf基因在结构上的保守性。同源性分析表明,黄喉拟水龟的Tf基因与鸟类、爬行类的Tf基因同源性最高,约为75%—97%;与哺乳类Tf基因的同源性约为65%—75%;与爪蟾等两栖类的Tf基因也有一定的同源性。荧光定量PCR结果显示,黄喉拟水龟人工感染粘质沙雷氏菌后,Tf基因在其肝脏、脾脏、肾脏及心脏各组织中的表达量均有上调的趋势,这与其他动物经病原刺激后的表达特征具有相似性。

     

    Abstract: Transferrin plays an important role in the iron metabolism and participates in the immune response after infectedby the bacterial. The full-length Tf genomic cDNA sequence was cloned, and the characteristics of Tf gene sequencewere analyzed. Sequencing analysis showed that Tf of the Yellow pond turtle constituted a single peptide chainby two similar domains, each domain consisted of two subunits, which interacted each other and formed a deep, hydrophiliciron binding sites. The Tf genomic DNA contained 17 exons and 16 introns, and the structure of Tf gene similar toother vertebrates, which showed that the Tf gene structure of the Yellow Pond Turtle was conserved. Homologousanalysis showed that the homology of the Tf gene among the Yellow Pond Turtle, birds and reptiles was about75%—97%; the homology of the Tf gene between the Yellow Pond Turtle and mammalian 65%—75%; the homology ofthe Tf gene among the Yellow Pond Turtle, Xenopus laevis and other amphibians was very little. Quantitative PCR resultsshowed that the expression of MaTf in the liver, spleen, kidney and heart tissues increased after infected by Serratiamarcescens in different time, and the characteristics was similar with other animals which were infected by a pathogen.

     

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