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热应激下虹鳟mRNA差异表达及分子调控机理研究

热应激下虹鳟mRNA差异表达及分子调控机理研究
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  • 批准号:31460687
  • 批准年度: 2014年
  • 学科分类:水产生物生理学(C190101) |
  • 项目负责人:刘哲
  • 负责人职称:教授
  • 依托单位:甘肃农业大学
  • 资助金额:52万元
  • 项目类别:地区科学基金项目
  • 研究期限:2015年01月01日 至 2018年12月31日
  • 中文关键词: 应激;虹鳟;mRNA;差异
  • 英文关键词:rainbow trout;RNA-Seq;heat stress;regulatory mechanism

项目摘要

中文摘要

虹鳟是我国主要的淡水冷水性养殖鱼类,但对高温耐受性差严重制约着其养殖业发展。目前,对应答虹鳟热应激的基因及其调控机理仍不清楚。项目以全雌虹鳟全同胞家系为材料,首先研究不同水温下虹鳟肝损伤、血细胞凋亡、血清免疫和代谢指标变化情况,评判不同水温下虹鳟的热应激反应(HSR)程度,确定HSR变化显著的"关键高温点"。然后基于高通量测序平台HiSeq 2500,进行RNA-Seq,构建虹鳟肝脏和肾脏"关键高温点"和最适水温(18℃)两组间差异基因表达谱,筛选出高温应激前后表达差异显著的蛋白编码基因,对其进行GO分析和KEGG通路分析,并对差异大于200倍的基因通过Real-time PCR进行验证,进而选择HSR关键信号通路上的核心基因3-5个,研究其在HSR过程中,在虹鳟6个组织(血液、肝、肾、脑、鳃和脾)中的时空表达差异,最终揭示虹鳟HSR分子调控机理,为虹鳟耐高温品种选育和抗热应激技术奠定基础。

英文摘要

Rainbow trout (Oncorhynchus mykiss.) is a cold-water species of salmonid, which is cultivated as the main varieties in our country, but its poor resistance to heat seriously restricts the development of aquaculture. So far, the genes involved in heat-shock and their molecular regulation mechanisms still remain unclear in rainbow trout. This project will firstly launch studies on status of liver damage, blood cell apoptosis, serum immune and changes of metabolism parameters under varied temperatures by utilization of all-female full-sib fishes, and the "key high-temperature point" (HTP) indicates the significant changes of heat stress response (HSR) will be defined. Then the differential gene expression profiles of liver and kidney between 18℃ and HTP will be set up respectively through RNA-Seq base on the next-generation sequencing system of HiSeq 2500, and the protein-coding genes differentially expressed will be screened and will be entered into Gene Ontology (GO) and KEGG pathway analysis. Furthermore, the genes of expression differences more than 200 times between 18℃ and HTP will be verified through the Real-time quantitative PCR, and three to five important genes in key HSR pathway will further functionally analyzed by investigating their temporal and spatial expression patterns in six different tissues (blood,liver, kidney, brain, spleen and gills). These results will uncover the molecular mechanisms of HSR,and will contribute to the development of heat resistant breeding programs and heat resistant technology in rainbow trout.

评估说明

    国家自然科学基金项目“热应激下虹鳟mRNA差异表达及分子调控机理研究”发布于爱科学iikx,并永久归类于相关科学基金导航中,仅供广大科研工作者查询、学习、选题参考。国科金是根据国家发展科学技术的方针、政策和规划,以及科学技术发展方向,面向全国资助基础研究和应用研究,发挥着促进我国基础研究源头创新的作用。国科金的真正价值在于它能否为科学进步和社会发展带来积极的影响。

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