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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (20): 148-153.doi: 10.11924/j.issn.1000-6850.2012-0859

所属专题： 生物技术

• 水产 渔业 • 上一篇    下一篇

虹鳟Hepcidin cDNA的克隆及序列分析

姜娜 马志宏 侯莉 邢薇 李铁梁 罗琳   

  • 收稿日期:2012-03-12 修回日期:2012-04-18 出版日期:2012-07-15 发布日期:2012-07-15
  • 基金资助:

    鱼用抗菌肽制备与应用关键技术研究

Cloning and Sequence Analysis of Antimicrobial Peptide Hepcidin from Rainbow Trout (Oncorhynchus mykiss)

  • Received:2012-03-12 Revised:2012-04-18 Online:2012-07-15 Published:2012-07-15

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摘要：

为了从虹鳟肝脏中扩增出Hepcidinc全长DNA序列，预测出氨基酸序列，并进行序列分析。通过RT-PCR和RACE PCR的方法，从虹鳟肝脏中克隆获得了Hepcidin 全长cDNA序列，利用在线工具和软件包分析。结果表明：虹鳟Hepcidin全长cDNA序列（GenBank登录号HQ711993）为883 bp，5’端非翻译区211 bp，3’端非翻译区为405 bp，以及一段267 bp的编码序列，编码88个氨基酸，由信号肽（24个氨基酸）、原肽（39个氨基酸）和成熟肽（25个氨基酸）组成，成熟肽C端含有Hepcidin类抗菌肽的8个半胱氨酸保守性结构，可形成4个分子内二硫键。与已报道的鲑形目大西洋鲑Hepcidin氨基酸序列的一致性为93%，与其他鱼类Hepcidin氨基酸序列的一致性为44%~88%，是Hepcidin家族的新成员。

关键词: 拯救CIAV M突变株, 拯救CIAV M突变株, 宿主细胞MSB1, 生长周期, 凋亡

Abstract:

The cDNA sequence of Hepcidin (hepatic bactericidal protein), an antimicrobial peptide and iron regulatory molecule primarily expressed in liver, were cloned and analysed. A full-length cDNA sequence of Hepcidin from rainbow trout (GenBank accession number: HQ711993) was obtained by RT-PCR and RACE and then analysed by bioinformatics software. The results showed that the length of Hepcidin cDNA was 883 bp with a CDS ranging from 212th to 475th bp, which encoded a peptide of 88 amino acids. The preprotide contained a signal peptide (24 amino acids), a prodomain (39 amino acids) and a mature peptide (25 amino acids). The 25-residue Hepcidin mature peptide included 8conserved cyseine residues which were predicted to form 4 intramolecular disulfide bonds. The deduced amino acid sequence of Hepcidin from rainbow trout shared 93% identity with those from Salmo salar and 44%~88% identity with those from other fishes. Hepcidin cDNA from Oncorhynchus mykiss was a new member of the Hepcidin gene family.