奥利亚罗非鱼DMO cDNA的分离和克隆

Cloning DMO cDNA of Oreochromis aurea

采用RTPCR和RACE法分离和测定了奥利亚罗非鱼DMOcDNA的全序列。得到1571bp[不含poly(A)]的全长cDNA,包括148bp5’非翻译区,1230bp阅读框以及含Poly(A)信号AATAAA的193bp3’非翻译区[不包括Poly(A)]。阅读框共编码409氨基酸,与尼罗罗非鱼DMO编码的氨基酸序列进行比较,同源性为96.3%,表明DMO在同一物种中差别较小。而与尼罗罗非鱼,红鳍东方豚,虹鳟,青鱼将,鼠,人等动物的DMRT1编码的氨基酸序列进行比较,同源性分别为:25.7%,25.8%,24.3%,29.7%,22.5%,22.0%,这说明DMO和DMRT1可能是两个不同的基因。

RT-PCR and RACE(rapid amplification cDNA ends ) were used for the isolation of the full length cDNA of DMO gene from ovary of Oreochromis aurea. Sequence analysis revealed a 1571 bp cDNA containing the 148 bp 5'-untranslated region, 193 bp 3'-untranslated region and 1230 bp open reading frame encoding 409 amino acid. Sequence analysis revealed the identity rate of deduced amino acid of DMO in O. aurea and O.niloticus is 96.3%, which showed high homology in species. However, we compared the alignment of deduced amino acid sequences between DMO cDNA from O.aurea and DMRT1 cDNA from O.niloticus, fugu, rainbow trout, medaka, rat and human. The score was 25.7%, 25.8%, 24.3%, 29.7%, 22.5% and 22.0%,respectively. These results indicated that DMO and DMRT1 gene may be different genes.