斜带石斑鱼抗菌肽hepcidin基因克隆及其成熟肽的原核融合表达

文章根据已报道的硬骨鱼类hepcidin cDNA序列设计简并引物，通过RT—PCR从重要海水经济鱼类斜带石斑鱼（Epinephelus coioides）肝脏中扩增出1条具有完整开放阅读框的246bpcDNA序列，Blast分析表明该序列是鱼类hepcidin基因家族的一员，被命名为斜带石斑鱼hepcidin样抗菌肽前体，该cDNA所推导的氨基酸序列有如下特点：1）信号肽序列与多数鱼类hepcidin信号肽的同源性在67％～87％之间；2）C端20个氨基酸序列具有绝大多数动物hepcidin成熟肽的共同典型保守序列特征，即在对应位置具有8个Cys；3）序列中不具有已报道的hepcidin前体肽转化酶作用典型基序RX（K／R）R]；4）与GenBank中已注册的2条斜带石斑鱼hepcidincDNA序列（GU391241和GU391242）所推导的氨基酸序列同源性分别为36％和33％，且NJ进化树显示不与这2条序列聚为一枝，表明该序列是斜带石斑鱼hepcidin基因家族中一种新亚型，其预测成熟肽融合表达载体成功在大肠杆菌（Escherichiacoli）中表达，为后续研究奠定基础。

Cloning of antimicrobial peptide hepcidin from Epinephelus coioides and prokaryotic fusion expression of its predicted mature peptide

Based on the known hepcidin cDNA sequences in teleosts,we designed some degenerate primers and cloned a 246 bp cDNA sequence with complete open reading frame from the liver of Epinephelus coioides,an important commercial marine fish.According to Blast analysis,the obtained sequence,named as E.coioides hepcidin-like antimicrobial peptide precursor,is a member of fish hepcidin gene family.The deduced amino acid sequence has such features as follows: 1) the similarity of hepcidins between the present signal pe...