广西眼镜蛇Dmrt基因DM结构域克隆及序列分析

Cloning and sequence analysis of DM domain of Dmrt gene in Guangxi Naja naja atra

【目的】了解广西眼镜蛇Dmrt基因DM结构域特性,为探讨眼镜蛇性别决定和分化发育的分子机制及证实Dmrt基因家族在爬行动物进化中的保守性提供理论依据。【方法】以广西眼镜蛇为材料,采用简并PCR扩增克隆眼镜蛇Dmrt基因DM结构域,并分析其与人、猕猴、牛、小鼠、大绿蛙、中华蟾蜍、扬子鳄等相应Dmrt基因DM结构域的同源性及系统进化关系。【结果】克隆获得广西眼镜蛇Dmrt基因家族的两个成员,分别命名为Nn1 Dmrt和Nn2 Dmrt。经系统进化分析,发现两个Dmrt基因DM保守区核酸序列与人、猕猴、牛、小鼠、大绿蛙、中华蟾蜍、扬子鳄相应Dmrt基因保守区核酸序列的相似性均在74%以上,其推导氨基酸序列的相似性均在80%以上。【结论】Dmrt基因DM结构域编码序列在不同物种中同源性较高,Dmrt基因家族在动物系统进化上具有高度的保守性。

[Objective]The present experiment was conducted to study the features of DM domain of Dmrt gene in order to explore the molecular mechanism of sex determination and differential development of Naja naja atra,and to explore the conservation of Dmrt gene in reptile evolution.[Method]Degenerate PCR was used to amplify DM domains of Dmrt gene in Naja naja atra.Its homology and evolutionary relationship with people,macaque,cow,mouse,Rana livida,Asiatic toad,and Chinese alligator were assayed.[Result]Two DM members were identified and named as Nn1 Dmrt and Nn2 Dmrt.The results of systemic evolutionary analysis indicated that nucleic acid sequences of these two DM members in the above-mentioned species shared a high identity（74%） and their amino acid sequences also shared a high similarity（80%）.[Conclusion]DM domain encoding sequence of Dmrt gene has high homology to various species,and also,the Dmrt gene family was immensely conserved during systemic evolution.