目录
摘要Ⅲ
关键词Ⅲ
AbstractⅣ
引言
引言
1材料与方法1
1.1材料 1
1.1.1植物材料1
1.1.2菌株与载体1
1.2实验方法1
1.2.1质粒的构建1
1.2.2酵母文库的筛选1
1.2.3验证4
2结果与分析4
2.1诱饵蛋白自激活检测结果4
2.2总酵母蛋白的提取结果5
2.3诱饵蛋白在酵母中western blot检测结果 5
2.4酵母文库的筛选结果5
2.5酵母质粒的提取结果6
2.6酵母质粒的测序结果6
3讨论7
3.1与ROS1互作的蛋白7
3.2延伸7
致谢7
参考文献8
拟南芥DNA去甲基化酶ROS1互作蛋白的筛选
摘要
DNA甲基化修饰对生物体有重要意义，它在塑造染色质结构、沉默转座元件和调控基因表达等方面都发挥着重要作用。DNA甲基化与去甲基化是表观遗传学的重要标志，植物在DNA甲基化和去甲基化的协调作用下维持基因组甲基化水平的动态平衡。DNA甲基化可以通过改变染色质的结构、DNA构象、组蛋白修饰及DNA与蛋白质间的互作方式对基因的表达进行调控，而DNA去甲基化是一个复杂的过程，可能涉及多个蛋白因子，对于DNA去甲基化的过程仍需深入探索。ROS1( repressor of silencing 1 )是一个重要的DNA去甲基化酶，它能够去除DNA上的甲基化修饰，对于维持转基因和内源基因的表达有重要意义。植物体内DNA甲基化的建立主要是通过RdDM途径，而ROS1能够反作用于RdDM途径的靶位点，进行DNA去甲基化的作用。研究ROS1的作用机制能使我们对植物DNA去甲基化过程有更深入的认识，本实验利用酵母双杂交系统，以ROS1蛋白为诱饵，筛选其互作蛋白，为深入解析DNA去甲基化的分子机理提供新的认识。
SCREENING OF INTERACTING PROTEIN OF ROS1 DNA DEMETHYLASE *51今日免费论文网|www.51jrft.com +Q: ￥351916072$ 
IN ARABIDOPSIS THALIANA
ABSTRACT
DNA methylation plays an important role in silencing transposable elements, shaping chromatin structure and regulating gene expression. DNA methylation is an important epigenetic marks. Plants maintain the dynamic balance of genome methylation level under the joint action of DNA methylation and demethylation. DNA methylation can regulate gene expression by changing chromatin structure, DNA conformation, histone modification, and DNAprotein interaction. DNA demethylation is a complex process that may involve multiple protein factors, and the process of DNA demethylation remains to be further understood. ROS1 ( repressor of silencing 1 ) is an important DNA demethylase, and ROS1 is a bifunctional enzyme. So ROS1 is important to maintain the expression of transgenic and homologous endogenous genes. DNA methylation in plants is established primarily through the RdDM pathway; actually, ROS1 can be guided to the target sites of RdDM pathway to conduct DNA demethylation. Therefore, investigations of the action mechanism of ROS1 is helpful for us to further recognize the demethylation process of plant DNA. In this study, the yeast twohybrid system was used, and ROS1 protein was used as bait to screen its interacting proteins. The discovered ROS1interacting proteins will facilitate the further understanding of molecular mechanisms underlying DNA demethylation.
KEY WORDS: DNA demethylation; ROS1; yeast twohybrid technology; protein interaction

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