Ran基因对中华鳖雌性性别分化的调控作用及机制研究

性别决定与分化是生命科学的热点和难点，是水产性控育种的关键问题。中华鳖兼具科研与经济价值，其生长存在两性异形，单性育种是产业亟待解决的关键技术，但迄今学术界对其性别决定和分化机理的了解还非常有限。本团队通过中华鳖基因组学和转录组学联合分析，鉴定出一批性别特异基因，其中的Ran在卵巢组织中特异表达。Ran是参与细胞分裂的重要调控因子，其功能在真核生物间具有高度保守性。然而，还未有关于Ran调控机体性别分化的研究报道。本项目拟定以中华鳖为研究对象，探索Ran对动物性别分化的作用：通过DNA-FISH，定位Ran于染色体；而后通过qPCR和原位杂交，分析Ran的时空表达模式及细胞定位，确定其作用的靶细胞；进而通过基因缺失和过表达，在体内、外分析Ran对中华鳖性别分化和卵巢发育的作用及调控机制。将为解析龟鳖动物性别分化机制提供新的线索，更为进一步探索Ran在脊椎动物生殖发育过程中的新功能奠定基础。

For a long time, the sex determination and differentiation in vertebrates has been a hot topic and also an important issue in sexual control breeding in aquaculture. In aquaculture, the males are more popular because their growth rate are faster than females. Moreover, the studies on all-male Chinese soft-shelled turtle have attracted more attention in aquaculture. However, the knowledge on sex determination and identification in turtle is still limited. In our previous work, we have screened some female specific genes through combining the genomics and transcriptomics analysis. One of the selected genes was Ran gene, which is exclusively expressed in the ovary in turtle. Ran is an important regulatory factor involved in cell division, and has highly conserved functions in eukaryotes, while there is lack of study on the function of Ran gene in sex differentiation. In this study, firstly, we will determine chromosomal location of Ran by DNA-FISH. Secondly, we will analyze the temporal-spatial expression and cellular localization of Ran gene by qPCR and in situ hybridization. Thirdly, we will demonstrate the regulation of Ran gene on sex differentiation and ovary development through loss and gain of gene function assay. The findings of this study will provide new clues for further investigations on sex differentiation in turtles. It would also be a prelude for exploring the new functions of Ran gene in vertebrates.