尼罗罗非鱼2个Dmrt基因的序列分析

Sequence Analysis of Dmrt Genes in Oreochromis niloticus

参照不同物种DM结构域设计了一对兼并引物,扩增出一条带,长约150bp。对扩增产物进行克隆,阳性克隆采用SSCP分析筛选不同基因片段,进一步对筛选的不同基因进行了DNA测序。结果获得2个不同的Dmrt基因,采用BLAST方法与人类相应Dmrt基因进行序列比对,发现与人类相应Dmrt基因的氨基酸序列一致性分别为89.1%和100%。根据尼罗罗非鱼(Oreochromis niloticus)的拉丁文名,按习惯将其命名为onDmrt1,onDmrt2。与其他动物相关的Dmrt基因进行聚类分析,结果表明,不同进化地位动物的Dmrt基因DM域编码序列存在高度的同源性,显示Dmrt基因在系统进化上高度保守,序列上的相似性可能暗示着它们在功能上的保守性。

The Dmrt genes of O. niloticus were amplified by using a pair of degenerated primers which were designed based on the conservative DM domain of different species. The amplification band of Dmrt gene familay was observed whose length was 150bp. Then, PCR products were cloned. To detect the different clones, SSCP technique was used. Two different Dmrt genes were obtained and sequenced respectively. After sequence analysis, those two Dmrt genes were named as on Dmrt1, onDmrt2 according to their high homology to corresponding human Dmrt genes because their identities to those human Dmrt genes in the amino acid sequence were 89.1%, 100% respectively; This may be concluded that Dmrt genes were highly conserved in phylogenetic.