杜氏盐藻PKC在盐胁迫信号传导中的功能研究

项目名称： 杜氏盐藻PKC在盐胁迫信号传导中的功能研究

项目编号： No.31472260

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： 柴晓杰

作者单位： 大连海洋大学

项目金额： 85万元

中文摘要： 本项目针对盐藻蛋白激酶C（Protein kinase C，PKC)信号途径在盐胁迫下的分子响应，拟采用分子生物学、蛋白质组学、计算生物学和生物信息学等多学科交叉的手段，系统研究盐藻PKC在抗盐胁迫中的功能。采用RT-PCR、RACE及实时荧光定量PCR技术克隆PKC基因，分析基因表达模式及其调控规律；RNAi技术鉴定PKC基因的功能；融合报告基因定位法确定PKC的亚细胞定位及移位情况；应用免疫共沉淀、酵母双杂交以及蛋白质组学的方法筛选PKC信号调控网络中的互作蛋白；利用分子对接和分子动力学模拟方法研究PKC与靶分子间的调控模式，构建盐藻PKC信号调控网络。本项目对细胞内典型的信号转导通路PKC途径进行系统深入的研究，探讨PKC基因及其信号调控网络在盐藻抗逆胁迫中的作用，从蛋白质组学水平揭示盐藻抗逆防御的分子机理，为阐明盐藻耐盐的分子机制提供新思路和新途径，有重要科学意义和经济利用价值。

中文关键词： 杜氏盐藻；PKC；信号通路网络；分子动力学；蛋白质组学

英文摘要： The research is carried out through the molecular response mechanism of Protein kinase C（PKC）signal pathway in Dunaliella salina induced by salt stress. Combining cross-disciplines method, molecular biology, proteome, computational biology and bioinformatics, will be used to systemically study the function of PKC in response to salt stress. The clone and functional analysis of PKC genes will be obtained using RT-PCR, RACE, QRT-PCR, and RNAi.The subcellular location and translocation of PKC proteins will also be investigated using fusion report gene mapping method.The interacting proteins in PKC signal regulation network will be screened by Co-immunoprecipitation assay and Yeast Two Hybrid System.Using simulation methods of bioinformatics, molecule linkage and molecule dynamics,we will reveal their interacting pattern.Finally the signal transduction by PKC pathway and protein expression controlling mechanisms to high salinity could be revealed by signal pathway network.The research focuses on location and shift of PKC related proteins in signal transduction pathway, interaction between these proteins and signal network. The aim of our research is to deeply illuminate molecular mechanism of PKC signal pathway of Dunaliella salina induced by salt stress. It is important to reveal the role of PKC gene and PKC related signal pathway network of Dunaliella salina induced by salt stress at the proteomics level. This will provide new thinking and new approach for the study of molecular mechanism of Dunaliella salina in response to stress. This study has both scientific significance and application values.

英文关键词： Dunaliella salina;PKC;Signal pathway network;Molecular dynamics;Proteomics