摘要
Dmrt(Doble-sex and mab-3 relatated transcription)基因家族是一系列广泛存在动物界的发育转录因子,参与生物体性别决定与分化、早期胚胎发育及器官组织功能稳定的维持等过程。本研究克隆和鉴别了三角帆蚌(Hyriopsis cumingii)中Dmrt家族的亚族Dmrta的基因,与Dmrta2同源性很高,将其命名为HcDmrta2-1。该基因全长1723 bp(登录号:MK372349),开放阅读框1158 bp,编码385个氨基酸,5’非编码区为237 bp,3’非编码区为328 bp。预测理论等电点/蛋白相对分子量为4.97/143293.27,不含信号肽或跨膜结构,存在1个DM保守结构域和1个DMA保守基序。采用荧光定量和原位杂交技术鉴定该基因在各组织及发育不同时期性腺中的表达及定位分析。结果表明,HcDmrta2-1基因在2龄成熟蚌的性腺、鳃、肝脏、肾脏、外套膜、闭壳肌及斧足中均表达,除了肝脏,其它各组织中雌雄表达差异表现为极显著(p<0.01),其中雌性性腺表达明显高于雄性;早期不同发育阶段的性腺表达在1月龄出现最高值,8月龄表达量次之,推测HcDmrta2-1参与了三角帆蚌幼龄性腺发育过程。2龄性腺组织原位杂交结果表明在雌雄个体中均检测到染色信号,且在雌性中很强,雄性中较弱。由此推测HcDmrta2-1基因参与了三角帆蚌幼龄发育和分化过程,且对雌蚌性腺的分化和成熟雄蚌性腺的功能维持起到重要作用。本研究对探讨淡水贝类中Dmrt家族的保守性及其性别决定机制和功能探索具有借鉴意义,为农业生产和人工养殖提供理论指导。
The Dmrt(Doble-sex and mab-3 relatated transcription)gene family is a series of developmental transcription factors widely present in the animal kingdom,involved in the process of sex determination and differentiation,early embryonic development,and function maintenance of organ or tissue.This study cloned and identified the gene from subfamily of Dmrt family in Hyriopsis cumingii,which had high homology with Dmrta2 and named HcDmrta2-1.Full length of HcDmrta2-1 was 1723 bp(Accession number:MK372349),containing an open reading frame of 1158 bp,encoding 385 amino acids,a 5’untranslated region of 237 bp and a 3’untranslated region of 328 bp.The predicted theoretical isoelectric point/protein relative molecular weight was 4.97/143293.27,without signal peptide or transmembrane structure,and existed one conserved DM domain and one DMA motif.Fluorescence quantification and in situ hybridization were used to identify the expression and localization of HcDmrta2-1 in various tissues and developmental genomic glands.The results showed that the HcDmrta2-1 gene was expressed in the gonads,sputum,liver,kidney,mantle,adductor and foot of 2-year-old individual.Except for the liver,the expression of male and female in other tissues was extremely significant(p<0.01),in which the expression of female gonads was extremely higher than that of males;Early expression of gonads at different developmental stages showed the highest level at 1 month,followed by 8 months,predicting that HcDmrta2-1 was involved in the development of larvae and gonada.In situ hybridization of 2-year-old gonad indicated that the staining signal was detected in both male and female individuals,and stronger in females and weaker in males.It is speculated that HcDmrta2-1 was related to the process of juvenile development and gonadal differentiation,and played an important role in the differentiation of female gonads and the maintenance of mature male gonads.This study is useful for exploring the conservation of the Dmrt family in freshwater shellfish and its
作者
葛静远
汪桂玲
吴丛迪
程起超
李家乐
Ge Jingyuan;Wang Guiling;Wu Congdi;Cheng Qichao;Li Jiale(Key Laboratory of Genetic Resources for Freshwater Aquaculture and Fisheries,Shanghai Ocean University,Shanghai 201306;Shanghai Engineer ing Research Center of Aquaculture,Shanghai,201306;National Demonstration Center for Experimental Fisheries Science Education,Shangha Ocean University,Shanghai,201306)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2020年第11期4992-5000,共9页
Genomics and Applied Biology
基金
国家重点研发计划“蓝色粮仓科技创新”专项计划资助(2018YFD0901400)
国家自然科学基金项目(3177-2835)
现代农业产业技术体系建设专项资金(CARS-49)共同资助。
