The study on molecular functions of the novel RNAi factors DRH-3 and E1 which are required to maintain the chromosome integrity in higher eukaryote

本研究旨在阐明我最初发现的线虫生殖系形成所必需的两个RNAi因子DRH-3(Dice Related Helicase-3)和E1(一种具有两个Tudor结构域的新蛋白)的分子功能。在这项研究中，我构建了10个突变蛋白的DNA表达载体，并利用大肠杆菌蛋白表达系统成功地制备了His和GST标记的DRH-3和E1蛋白的天然和部分突变版本，并用不同标记的天然蛋白通过下拉方法展示了DRH-3和E1之间的物理相互作用。此外，对这两个基因的遗传学和细胞学观察表明，DRH-3和E1蛋白在细胞核内通过相同的RNAi途径发挥作用。

This study has aimed to clarify the molecular function of two RNAi factors DRH-3 (Dicer Related Helicase-3) and E1 (a novel protein with two Tudor domains) that I originally found to be indispensable for germ-line formation in C. elegans. In this study, I generated the DNA constructs for expression of 10 mutated proteins and successfully prepared His- and GST-tagged-native and some of mutated versions of DRH-3 and E1 proteins using the E. coli protein expression system, and showed the physical interaction between DRH-3 and E1 by pull-down method with differently tagged-native proteins. In addition, genetic and cytological observations on both genes have suggested that DRH-3 and E1 proteins act in the same pathway of RNAi in nucleus.