根据其他鱼类DMRT基因中的保守序列设计了一对简并引物,利用RT-PCR扩增了雌雄奥利亚罗非鱼的DMRT基因,并对其扩增产物进行了克隆与测序。结果在雌雄奥利亚罗非鱼个体中获得了两个不同的片段,分别命名为DMO,DMRT1基因。序列同源性分析表明,奥利亚罗非鱼DMRT1,DMOcNA系列的同源性为61.78%,氨基酸同源性为86%,说明了DMRT基因存在性别差异。与尼罗罗非鱼、红鳍东方豚、虹鳟、青鳞等鱼类DM保守区相比,氨基酸序列同源性为86%-100%,这充分显示了DMRT基因在进化上的高度保守性。
Using a pair of degenerate primes, We amplified two 158bp cDNA fragments of the dnrt gene in filapia, Oreochrom/s aurea trom testis and ovary by RT- PCR. The sequences from male and fenalle tilapia were different. We named them dmrtl, and dmo gene respectively. The sequence analysis indicated that the eDNA sequence and the amino acids homogeneity between dmrtl and dmo gent were 61.78% and 86% respectively, which showed that dmrt gene was different in male and femal tilapia. We compared the aligmnent of deduced amino acid sequences between DMO eDNA from O. aurea, O nilotieus and DMRT1 cDNA from O. niloticus, fugu, rainbow trout, medaka. Tile score was 86% - 100%. These results indicated that DMO and DMRT1 genes were highly conservative in phylogeny.