唐永凯,李建林,李红霞,陈雪峰,俞菊华.2008.奥利亚罗非鱼EF-1α基因的克隆与结构分析.动物学杂志,43(5):7-12. |
奥利亚罗非鱼EF-1α基因的克隆与结构分析 |
Cloning and Gene Construction Analysis of EF-1α in Oreochromis aureus |
投稿时间:2008-03-13 修订日期:2008-06-24 |
DOI: |
中文关键词: 奥利亚罗非鱼 克隆 EF-1α |
英文关键词:Oreochromis aureus Clone EF-1α |
基金项目:中央级基本科研业务费专项项目(No.2007JBFC03);江苏省自然科学基金项目(No.BK2006029) |
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中文摘要: |
真核生物延伸生长因子基因(EF-1α)在蛋白质翻译过程中起着重要的作用,其序列具有高度的保守性,是一种管家基因。本文通过RT-PCR克隆出奥利亚罗非鱼(Oreochromis aureus)EF-1α的部分cDNA序列,其长度为425bp,翻译成141个氨基酸,计算的蛋白质分子量为15.1ku。同源性分析显示,奥利亚罗非鱼EF-1α氨基酸序列与尼罗罗非鱼(O.niloticus)的相似性最高,为100%;与青(鱼将)(Oryzias latipes)、欧洲鲈(Dicentrarchus labrax)、斑马鱼(Danio rerio)、鲑鱼(Salmo trutta)的相似性分别为92%、91%、85%、82%;与小鼠(Mus musculus)、大鼠(Rattus norvegicus)、人(Homo sapiens)、鸡(Gallus gallus)的相似性均为85%。同时克隆出奥利亚罗非鱼EF-1α相应的DNA序列,共506bp。cDNA与DNA的序列比对显示克隆出的奥利亚罗非鱼EF-1α含有1个内含子,这为将来设计EF-1α荧光定量引物以及测定其在不同组织中的表达量变化打下基础。 |
英文摘要: |
Eukaryotic elongation factor 1α(EF-1α)plays an important role in translation and its sequence is highly conservative as a housekeeping gene.The partial cDNA encoding EF-1α in Oreochromis aureus was isolated using RT-PCR.The cDNA encoding 141 amino acids with a calculated molecular weight of 15.1 ku was 425 bp.The comparison of the deduced amino acid sequence of O.aureus EF-1α with that of O.niloticus,Oryzias latipes,Dicentrarchus labrax,Danio rerio or Salmo trutta showed that the homology rates were 100%,92%,91%,85%,and 82%,respectively,while the amino acids homology was 85% when comparing O.aureus with Mus musculus,Homo sapiens,Gallus gallus,or Rattus norvegicus.The DNA sequence of O.aureus EF-1α consisting of 506 bp was also cloned.Comparing the partial cDNA to its genomic sequence revealed that O.aureus EF-1α gene consisted of an intron,which supplied useful information in designing the EF-1α primer in real time RT-PCR and calculating the EF-1α expression in different tissues. |
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