拟穴青蟹VIH的转录调控机制研究

项目名称： 拟穴青蟹VIH的转录调控机制研究

项目编号： No.31472266

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 王艺磊

作者单位： 集美大学

项目金额： 86万元

中文摘要： 由X-器官窦腺复合体组成的虾蟹眼柄内分泌系统对虾蟹生殖系统的发育和分化起着至关重要的调控作用。其合成和分泌的VIH抑制性腺发育。但是VIH基因在转录前如何被调控，到目前为止尚未见报道。在国家自然科学基金的资助下，课题组获得了拟穴青蟹VIH基因的全长cDNA和gDNA，通过Tail-PCR获得了转录起始位点上游部分序列。在此基础上，本项目将进一步延长拟穴青蟹VIH 5'上游调控区序列并分析其活性，确认VIH顺式作用元件的功能。同时鉴定调控VIH表达的重要转录因子，了解VIH表达与转录因子之间的功能相关性，确认转录因子对VIH的正负调控作用。通过ChIP-seq技术分析调控VIH的重要转录因子在X-器官窦腺复合体同步调控的其它性腺发育相关基因，阐明VIH的转录调控机制。研究成果将使我们在理论上更加深入了解拟穴青蟹性腺发育的调控机制，在生产实践为建立新的生殖调控技术提供新的思路。

中文关键词： VIH；顺式作用元件；转录因子；转录调控；拟穴青蟹

英文摘要： Members of crustacean hyperglycemic hormone family, including crustacean hyperglycemic hormone (CHH), molt inhibiting hormone (MIH), vitellogenesis inhibiting hormone (GIH), and mandibular organ inhibiting hormone (MOIH), are mostly secreted from the X-organ sinus gland complex of the eyestalk in crustacean. Similar to the hypothalamus-pituitary-gonad axis in vertebrates, the X-organ sinus gland complex and gonad in crustaceans form a similar axis to control the maturation of gonad. Many studies have confirmed that VIH inhibits vitellin synthesis in ovarian tissue. However, up to date, the mechanism of transcriptional regulation on expression of VIH gene have not been reported in crustaceans. Supported by the National Natural Science Foundation, VIH gene from Scylla paramamosain was cloned for the first time. The full length of cDNA, genomic DNA, and partial sequence of 5'-regulatory region of VIH of mud crab were obtained. Based on the progresses we have made in previous projects, in this project, 5' upstream regulatory region will be cloned and prolonged to 5-6 kb by Tail-PCR firstly. Secondly, the activity of 5' upstream regulatory region will be analyzed and the function of cis-element will be identified. Thirdly, the important transcriptional factors of VIH will be obtained, and the interact with cis-element will be identified. The functional correction between transcriptional factor and VIH mRNA will be elicited. Fourthly, Some gonad-related genes co-regulated by transcriptional factors of VIH in X-organ sinus gland complex will also be identified using ChIP-seq method. Finally, the mechanism of transcriptional regulation of VIH mRNA in S. paramamosain will be interpreted. The completion of this project will help us to demonstrate the molecular mechanism of transcriptioanl regulation of VIH in S. paramamosain. It will also help us to develop new reproduction regulation techniques for crab aquaculture.

英文关键词： VIH;cis-element;transcriptional factor;transcriptional regulation;Scylla paramamosain