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引用本文:葛建龙,陈晓辉,陈四清,刘长琳,陈昱辰,边力,张盛农.海蜇dmrta2基因的克隆及其在横裂生殖中的表达分析[J].海洋科学,2022,46(6):42-50.
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海蜇dmrta2基因的克隆及其在横裂生殖中的表达分析
葛建龙1, 陈晓辉1, 陈四清1, 刘长琳1, 陈昱辰2, 边力1, 张盛农1
1.中国水产科学研究院 黄海水产研究所, 青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266071;2.盐城市华亿水产有限公司, 江苏 盐城 224300
摘要:
Dmrt基因家族在后生动物性别决定与分化以及组织和器官的形成等发育过程中发挥着重要作用。为探究dmrt家族基因在海蜇(Rhopilema esculentum)横裂生殖中的功能,本研究以海蜇转录组测序获得的dmrta2基因片段为基础,通过RACE技术获得了海蜇dmrta2基因的cDNA全长,并分析了其在横裂生殖不同时期的表达模式。结果显示海蜇dmrta2基因cDNA全长为1 804 bp,开放阅读框为1 011 bp,所编码蛋白质含336个氨基酸,分子质量为37.25 kDa,理论等电点为9.11。预测海蜇dmrta2编码蛋白为亲水性蛋白质,无跨膜区域,不含信号肽。在33至79位氨基酸之间具有dmrt基因家族保守的DM结构域,与珊瑚、果蝇等物种的同源基因相应区域高度一致。系统进化分析显示,海蜇DMRTA2与鹿角珊瑚DMRTA2和DMRT3以及海葵DMRTA2的亲缘关系最近。原位杂交显示dmrta2基因在海蜇横裂生殖后期主要表达于感觉缘叶原基位置,在初生碟状体中表达于感觉棍位置。研究结果表明,dmrta2基因参与海蜇横裂生殖过程,可能主要与感觉棍神经系统的分化发育相关,研究为进一步解析海蜇等钵水母横裂生殖的分子调控机制提供了基础数据。
关键词:  海蜇  横裂生殖  Dmrta2  表达模式
DOI:10.11759/hykx20210308002
分类号:Q78
基金项目:政府间国际科技创新合作重点专项(2017YFE0111100-5);国家自然科学基金项目(31702327);农业农村部海洋渔业可持续发展重点实验室开放课题(20603022019020)
Cloning of dmrta2 from jellyfish Rhopilema esculentum and its expression analysis during strobilation
GE Jian-long1, CHEN Xiao-hui1, CHEN Si-qing1, LIU Chang-lin1, CHEN Yu-chen2, BIAN Li1, ZHANG Sheng-nong1
1.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Laboratory for Marine Fisheries Science and Food Production Process, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071, China;2.Huayi aquatic breeding limited company, Yancheng 224300, China
Abstract:
The dmrt gene family plays an important role in various developmental processes of metazoans, such as sex determination and differentiation and the formation of tissues and organs. Strobilation is a unique asexual reproductive mode of scyphozoan jellyfish and a key stage in the transformation from polyp to medusa. However, the understanding of molecular mechanisms under drastic metamorphosis is still limited. In a previous study, it was preliminary found that the dmrt gene family may play a crucial role in the strobilation of the jellyfish (Rhopilema esculentum); therefore, this finding is of great interest and needs further validation. In the present study, based on a dmrta2 fragment from the RNA-sequence of R. esculentum, full-length complementary DNA (cDNA) of dmrta2 was obtained using the rapid amplification of cDNA ends (RACE) method, and its expression patterns during strobilation were studied. The results showed that the full-length cDNA of R. esculentum dmrta2 was 1804 bp, including an open reading frame (ORF) of 1011 bp length, which encodes 336 amino acids. The complete sequence of R. esculentum dmrta2 has been submitted to the GenBank database with the following accession number: MT878494. The molecular weight (MW) and theoretical isoelectric point (PI) were predicted to be 37.25 kDa and 9.11, respectively. It was a hydrophilic protein comprising nontransmembrane structure and signal peptides. DMRTA2 of R. esculentum had a conserved DM-domain of dmrt gene family between the 33rd and 79th amino acid, which exhibits a great similarity to the homology protein in coral, fruit fly, and so on. Phylogenetic analysis showed that DMRTA2 of R. esculentum was closely related to the DMRTA2,DMRT3 of coral Acropora millepora and DMRTA2 of sea anemone Nematostella vectensis. The whole-mount in situ hybridizations showed that dmrta2 lacked expression signal in the polyp stage, whereas it was expressed in the tentacle base of the early strobila stage, expressed distinctly in the pre-rhopalar lappets of late strobila and in the rhopalium region of ephyra. The results indicated that dmrta2 was involved in the strobilation of R. esculentum and probably related to the differentiation and development of the rhopalium nervous system. To the best of our knowledge, all these results may provide important references for future studies of molecular regulation mechanisms of scyphozoan strobilation.
Key words:  Rhopilema esculentum  strobilation  dmrta2  expression pattern
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