黄鳝转铁蛋白受体1基因的克隆及表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2015.07.019

生物技术通报 ›› 2015, Vol. 31 ›› Issue (7): 132-137.doi: 10.13560/j.cnki.biotech.bull.1985.2015.07.019

黄鳝转铁蛋白受体1基因的克隆及表达分析

江翱¹, 李伟^1，2

(1.长江大学生命科学学院，荆州 434025；2.湿地生态与农业利用教育部工程研究中心，荆州 434025)

收稿日期:2014-11-12 出版日期:2015-07-16 发布日期:2015-07-16
作者简介:江翱，男，研究方向：生物技术制药；E-mail：1362541137@qq.com
基金资助:
湖北省教育厅基金项目(Q20131206)，湿地生态与农业利用教育部工程研究中心开放课题(2013A011)，国家大学生创新创业训练计划项目(104892014032)

Cloning and Expression Analysis of a Transferrin Receptor 1 Gene from Swamp Eel, Monopterus albus

Jiang Ao¹, Li Wei^1，2

(1. College of Life Science，Yangtze University，Jingzhou 434025；2. Engineering Research Center of Ecology and Agricultural Use of Wetland，Ministry of Education，Jingzhou 434025)

Received:2014-11-12 Published:2015-07-16 Online:2015-07-16

摘要/Abstract

摘要： 旨在研究黄鳝转铁蛋白受体1基因的功能及其组织表达特异性，采用同源克隆结合RACE技术从黄鳝肝脏中分离其转铁蛋白受体1基因，用生物信息学方法对获得的氨基酸序列进行分析，用半定量RT-PCR技术研究各组织中TfR1表达水平。结果表明，克隆获得黄鳝TfR1，GenBank注册序列号为KF819396。该cDNA全长2 839 bp，5' UTR长134 bp，3' UTR长380 bp，编码一个长774个氨基酸的多肽链。氨基酸多序列比对结果表明，黄鳝TfR1基因推断的氨基酸序列同其他鱼类的同源性较高，达55.68%-68.41%；而同哺乳动物的TfR1基因同源性较低。半定量RT-PCR分析表明，黄鳝TfR1基因转录本在不同组织表达量有明显差异；在血细胞中表达量最高，而在肾脏、脾脏和小肠中表达中等，在肝脏、胃、皮肤、脑、心脏和肌肉中表达量很低。

关键词: 黄鳝, 转铁蛋白受体1, RACE技术, RT-PCR技术

Abstract: For investigating the function and tissue expression specificity of transferrin receptor 1 gene(TfR1)from swamp eel, TfR1 gene was isolated from the liver of swamp eel using homologous cloning and RACE techniques, the bioinformatics method was used to analyze the putative amino acid sequence, and semi-quantitative RT-PCR was used to study the expression of TfR1 gene in different tissues. The results showed that the full length of cDNA(GenBank accession number：KF819396)was 2 839 bp including a 134 bp 5' un-translated region(UTR)and a 380 bp 3' UTR, and it encoded a polypeptide of 774 amino acids. Sequence alignment by amino acid indicated that the inferred amino acid sequence of TfR1 from swamp eel shared high identity with those of other teleosts in 55.68%-68.41%. Semi-quantitative RT-PCR analyses demonstrated that the TfR1 transcripts were significantly varied in different tissues, the highest in blood cells, moderate in kidney, spleen and intestine, and little in liver, stomach, skin, brain, heart and muscle.

Key words: swamp eel, transferrin receptor 1(TfR1), RACE, RT-PCR

江翱, 李伟. 黄鳝转铁蛋白受体1基因的克隆及表达分析[J]. 生物技术通报, 2015, 31(7): 132-137.

Jiang Ao, Li Wei. Cloning and Expression Analysis of a Transferrin Receptor 1 Gene from Swamp Eel, Monopterus albus[J]. Biotechnology Bulletin, 2015, 31(7): 132-137.

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黄鳝转铁蛋白受体1基因的克隆及表达分析

Cloning and Expression Analysis of a Transferrin Receptor 1 Gene from Swamp Eel, Monopterus albus

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