文章摘要
胡海彦,贾永义,唐永凯.奥利亚罗非鱼β-actin基因的克隆与结构分析[J].上海海洋大学学报,2011,20(2):185-190
奥利亚罗非鱼β-actin基因的克隆与结构分析
Cloning and gene construction analysis of β-actin in Oreochromis aureus
  
DOI:
中文关键词: 奥利亚罗非鱼  β-actin  克隆
英文关键词: Oreochromis aureus  β-actin  cloning
基金项目:现代农业产业技术体系建设专项资金(nycytx-49); 基本科研业务费专项资金(2011JBFC09)
作者单位
胡海彦 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 
贾永义 浙江省淡水水产研究所 
唐永凯 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 
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中文摘要:
      β-actin在真核细胞的生理过程中起着重要作用,其序列具有高度保守性,是一种管家基因。通过RT-PCR方法克隆出奥利亚罗非鱼(Oreochromis aureus)β-actin的部分cDNA序列,其长度为424 bp,翻译成138个氨基酸,计算的蛋白质分子量为15.5 ku。氨基酸同源性分析显示,奥利亚罗非鱼β-actin与真鲷(Pagrus major)、斑马鱼(Danio rerio)、青鳉(Oryzias latipes)、龙溪鳉(Rivulus marmoratus)的相似性最高,为99.3%;与鲫(Carassius auratus)、红鳍东方鲀(Takifugu rubripes)等其它鱼的相似性也较高,为97.8%~98.6%。此外,还克隆出了奥利亚罗非鱼β-actin〖WTBZ〗相应的DNA序列,共619 bp。cDNA和DNA的序列比对显示克隆出的奥利亚罗非鱼β-actin含有2个内含子,这为将来设计β actin〖WTBZ〗荧光定量PCR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 β-actin gene plays an important role in physiological process and its sequence is highly conservative as a housekeeping gene. The partial cDNA encoding β-actin in Oreochromis aureus was isolated using RT- PCR. The cDNA isolated was 424 bp encoding 138 amino acids with a calculated molecular weight of 15.5 ku. The comparison analysis of the deduced amino acid sequence of β-actin in O.aureus showed high similarity of 99.3% with those of Pagrus major, Danio rerio, Oryzias latipes, Rivulus marmoratus, and 97.8%-98.6% homology with those of other fishes, such as Carassius auratus, Takifugu rubripes. The corresponding DNA sequence of O.aureus β-actin in size of 619 bp was also cloned. By comparing the sequences of cDNA and DNA, it was found that β-actin gene in O.aureus consisted of two introns. The present study provided some useful information in designing the β-actin primers for real time RT-PCR and evaluating the β-actin expression level for different tissues.
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