三角帆蚌Dmrt1基因的分子特征和表达研究  

Molecular Characterization and Expression Analysis of Dmrt1 Gene in Hyriopsis cumingii

作　　者：郭鹏飞 段胜华 董赛赛 吴丛迪 汪桂玲[1] Guo Pengfei;Duan Shenhua;Dong Saisai;Wu Congdi;Wang Guiling(Shanghai Engineering Research Center of Aquaculture,Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources,National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai,201306)

机构地区：[1]上海海洋大学,水产科学国家实验教学示范中心,水产种质资源发掘与利用教育部重点实验室,上海市水产养殖工程技术研究中心,上海201306

出　　处：《基因组学与应用生物学》2020年第5期2033-2041,共9页Genomics and Applied Biology

基　　金：国家自然科学基金项目(31772835);现代农业产业技术体系建设专项资金(CARS-49)共同资助。

摘　　要：Dmrt(Doble-sex and mab-3 relatated transcription factor)基因的主要功能是决定性别、分化性腺和形成组织和器官。为研究其在三角帆蚌(Hyriopsis cumingii)性腺发育和分化过程中的作用,本实验通过比对三角帆蚌转录组序列和RACE扩增的方法,获得了三角帆蚌Dmrt1基因(HcDmrt1)全长cDNA,采用实时定量PCR的方法研究该基因在性腺发育和分化的表达情况。结果显示,HcDmrt1基因全长为1561 bp,ORF区(开放阅读框区域)1161 bp,编码386个氨基酸。编码蛋白具有DM(Doublesex/MAB-3)Domain family保守结构域。三角帆蚌2~10月龄个体性腺荧光定量结果显示该基因参与了早期性腺发育;组织荧光定量结果显示,HcDmrt1在各组织中均有表达,但是在雌雄性腺组织中表达差异最大,雄性中表达量明显高于雌性。性腺的原位杂交结果显示,该基因主要在卵母细胞的细胞膜上检测到表达信号,在雄性性腺中,主要在滤泡腔中的精细胞观察到信号。由此推测,HcDmrt1基因参与了三角帆蚌的性腺发育过程。The function of the Dmrt(Doble-sex and mab-3 relatated transcription factor)gene is mainly sex determination,gonadal differentiation and the formation of many tissues and organs.In order to study its role in the development and differentiation of the gonads of Hyriopsis cumingii,the full length of the Dmrt1 gene(HcDmrt1)was obtained by comparing the transcriptome sequence and the RACE amplification method of Hyriopsis cumingii cDNA,Real-time quantitative PCR was used to study the expression of this gene in gonadal development and differentiation.The results showed that the HcDmrt1 gene was 1561 bp in length and the ORF region(open reading frame region)was 1161 bp,encoding 386 amino acids.The encoded protein has a DM(Doublesex/MAB-3)Domain family conserved domain.The quantitative analysis of gonadal tissue from 2~10 month showed that the gene was involved in early gonadal development.The results of tissue Real-time quantitative PCR analysis showed that HcDmrt1 was expressed in all tissues,but the expression was the most different in male and female gland tissues,and the expression level in males was significantly higher than that in females.The in situ hybridization of the gonads showed that the gene mainly detected the expression signal on the cell membrane of the oocyte,and in the male gonad,the sperm cells in the follicular cavity observed the signal.It is speculated that the HcDmrt1 gene is involved in the gonadal development of the spiny.

关 键 词：三角帆蚌 DMRT1 性腺发育 原位杂交 

分 类 号：S917.4[农业科学—水产科学]