鲢鱼解偶联蛋白2全长cDNA序列的克隆及其组织表达
cDNA Sequence Cloning and Tissue Expression of Uncoupling Protein 2 of Silver Carp (Hypophthalmichthys molitrix)
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摘要: 从淡水食毒藻鱼类鲢鱼(Hypophthalmichthys molitrix)肝脏，通过简并引物克隆解偶联蛋白2(uncoupling protein 2，UCP2) cDNA核心序列，应用5′RACE和3′RACE技术分别扩增该序列的5′末端和3′末端序列，最后通过序列拼接获得鲢鱼肝脏UCP2 cDNA全序列。序列分析结果表明，鲢鱼肝脏UCP2 cDNA全长1 452 bp，其中5′-UTR长337 bp，3′-UTR长182 bp，编码区933 bp，编码310个氨基酸，推测的氨基酸序列包含线粒体内膜载体蛋白3个特征结构及解偶联蛋白(UCPs)的特征序列。对鲢鱼不同组织UCP2的表达调控研究发现，鲢鱼组织UCP2基因在肠道、肝脏、肌肉、脂肪组织均大量表达，而在脑组织表达量较低，这与鲢鱼体内微囊藻毒素在这几个组织的分布完全一致，表明UCP2的功能可能与抑制微囊藻毒素引发过量活性氧(ROS)生成有关。Abstract: PCR using two degenerated primers designed on the basis of the conserved region of mammals and fish uncoupling protein 2 (UCP2) gene yielded a cDNA fragment of 620 bp in the liver of silver carp. This cDNA fragment was completed by 5′ and 3′ RACE. The full-length silver carp UCP2 cDNA clone was 1 452 bp in length, containing an open reading frame of 933 bp (encoding 310 amino acids), flanked by 337 bp 5′UTR and 182 bp 3′UTR. The deduced amino acid sequence from this UCP2 cDNA fragment contains three signature motifs of mitochondrial transporter protein, and UCP-specific sequences. Tissue-expression analysis revealed that the silver carp UCP2 was highly expressed in intestine, liver, muscle and adipose tissue, but lower in brain tissue. This tissue expression pattern is consistent with the microcystins distribution among these tissues, suggesting that UCP2 expression level has a close relationship with the overproduced ROS which is induced by microcystins.
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