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。
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.
出处
《上海海洋大学学报》
CAS
CSCD
北大核心
2011年第2期185-190,共6页
Journal of Shanghai Ocean University
基金
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)
