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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。
β-actin plays an important role in the physiological process of eukaryotic cells. Its sequence is highly conserved and is a housekeeping gene. A partial cDNA sequence of β-actin from Oreochromis aureus was cloned by RT-PCR. It was 424 bp in length and translated into 138 amino acids. The calculated protein molecular weight was 15.5 ku. Amino acid homology analysis showed that β-actin of O. tilapia had the highest similarity with Pagrus major, Danio rerio, Oryzias latipes and Rivulus marmoratus, Was 99.3%. The similarity to other fish such as Carassius auratus and Takifugu rubripes was also high, ranging from 97.8% to 98.6%. In addition, the corresponding DNA sequence of β-actin in Oreochromis niloticus was cloned, a total of 619 bp. The sequence alignment of cDNA and DNA showed that the cloned β-actin of O. tilapia contained two introns, which could lay the foundation for the future design of β-actin fluorescence quantitative PCR primers and determination of their expression changes in different tissues basis.