华北农学报

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黄鳝AQP1 cDNA的克隆与表达分析

丁炜东, 曹丽萍, 曹哲明, 邴旭文   

  1. 中国水产科学研究院淡水渔业研究中心农业部水生动物遗传育种和养殖生物学重点开放实验室, 江苏 无锡 214081
  • 收稿日期:2012-09-10 出版日期:2012-12-31
  • 通讯作者: 邴旭文(1968 - ), 男, 江苏无锡人, 研究员, 硕士, 主要从事特种水产养殖与育种研究。
  • 作者简介:丁炜东(1977 - ), 男, 江苏如东人, 硕士, 主要从事鱼类遗传育种研究。
  • 基金资助:
    公益性行业科研专项-鳝鱼产业技术研究与试验示范(201003076);中央级公益性科研院所基本科研业务费专项资金(中国水产科学研究院淡水渔业研究中心)项目(2010JBFC07)

Molecular Cloning of Aquaporin1 cDNA and Multi-tissue mRNA Expression in Monopertus albus

DING Wei-dong, CAO Li-ping, CAO Zhe-ming, BING Xu-wen   

  1. Key Laboratory for Genetic Breeding of Aquatic Animals and Aquaculture Biology Certificated by the Ministry of Agriculture,Freshwater Fisheries Research Center,Chinese Academy of Fishery Sciences,Wuxi 214081,China
  • Received:2012-09-10 Published:2012-12-31

摘要: 采用RT-PCR和cDNA末端快速扩增法(RACE)分离、克隆出黄鳝水通道蛋白(Aquaporin 1,AQP1)基因全长cDNA序列,共1 134 bp,包括163 bp 5’非翻译区、789 bp阅读框以及182 bp 3’非翻译区,阅读框共编码263个氨基酸,分子量计算值为27.8 kDa,理论等电点为7.945。氨基酸组成中,正电荷残基(K,R)总数21个,负电荷残基(E,D)总数19个,极性氨基酸为115个,疏水性氨基酸60个。同源性分析表明,黄鳝与欧洲鳗的相似性高达87%,其次与底(鱼将)、鲷、犬、克氏鲤、猪、珍珠鸟、斑马鱼、半滑舌鳎、非洲肺鱼、蟾蜍、非洲瓜蟾的相似性分别为42.9%,61.2%,46.1%,42.9%,53.9%,47.9%,41.5%,40.1%,33.2%,30.8%,33.2%。生物信息学分析显示,黄鳝AQP1基因所编码的蛋白质以亲水性区域为主,具有丰富的B细胞抗原位点。定量分析结果表明,AQP1 mRNA主要在卵巢中高表达,其他组织中表达量较少。

关键词: 黄鳝, 卵巢, 水通道蛋白

Abstract: Monopertus albus Aquaporin 1 ( AQP1 ) cDNA was cloned to investing its structure, evolution, and expression in different tissues by RT-PCR and RACE method.The result shown that the AQP1 cDNA is composed of a 163 bp 5'UTR, 182 bp 3 'UTR and a 1 134 bp nuclotides with a 952 bp nucloetide open reading frame encoding a protein of 263 amino acids.Protein sequence alignment showed that the highest similarity with the European eel ( Monopertus albus) AQP1 (87% ) and the other is 42.9% , 61 .2% , 46.1% , 42.9% , 53 .9% , 47.9% , 41 .5% ,40.1 % , 33 .2% , 30.8% , 33 .2% .Bioinformatics analysis revealed that the protein whose calculated Molecular weight is 27.8 kDa and theoretical isoelectric point is 7.945 was hydrophilic in most region and had rich B cell antigencity positions.The survey about multiple tissue expression of AQP1 gene in eel revealed that the AQP1 mRNA is mainly expressed in ovary and the concentration of mRNA in other tissues is very small.

Key words: Monopertus albus, Ovary, Aquaporin 1

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引用本文

DING Wei-dong, CAO Li-ping, CAO Zhe-ming, BING Xu-wen. Molecular Cloning of Aquaporin1 cDNA and Multi-tissue mRNA Expression in Monopertus albus[J]. ACTA AGRICULTURAE BOREALI-SINICA, doi: 10.3969/j.issn.1000-7091.2012.z1.002.

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