RUI: Molecular and Genetic Analysis of Two Putative Rad51 Paralogs in Drosophila melanogaster

RUI：果蝇中两个假定的 Rad51 旁系同源物的分子和遗传分析

• 批准号: 0221892
• 负责人: Lisa Wrischnik
• 金额: $ 10.96万
• 依托单位: University of the Pacific
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2005-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0221892&HistoricalAwards=false
• 关键词: RUI; Molecular; Genetic; Analysis; Two

Using sequence information gathered from the Drosophila Genome Project and Celera/Applied Biosystems, the Drosophila DNA Repair project has identified several paralogs of the yeast Rad51 gene. These paralogs are thought to promote Rad51-dependent strand exchange during the process of homologous recombinational repair (HRR) of DNA damage in cells. An analysis of hamster cell mutations, coupled with targeted knockout experiments in chicken cells, has shown that these genes play an important role in providing resistance to ionizing radiation and chemical mutagens. The focus of this research project is the study of DmRad51D and DmXRCC2 and their role in mitotic homologous recombination repair in fruit flies. The long-term goals include the use of genetic screens to identify other genes that interact with DmRad51D and DmXRCC2 in the process of HRR. The objectives of the research include 1) confirming the functional role of these putative D. melanogaster Rad51 paralogs by assaying functional knockouts in Drosophila culture cells, 2) examining the protein-protein interactions among these genes, 3) examining the expression pattern of these genes in flies, and 4) using P-element excision to generate loss of function mutations in the DmXRCC2 gene. DNA contains the instructions that allow cells to function correctly. However, DNA is constantly damaged by chemicals in cells and by radiation such as UV or gamma rays. Cells with damaged DNA do not always work properly and aberrations can result. A large group of genes repairs damaged DNA. This project focuses on the roles of two related genes in DNA repair. While scientists have some understanding of how these genes work in the test tube or in single cells, they still do not know much about how they work in a whole organism. This project will use molecular and genetic tools to study the role of these DNA repair genes in the fruit fly, where the activity of these genes can be studied in the entire animal. Almost all of the genes that fruit flies use during growth and development play an important role in all animals. This project will not only use the fruit fly to explore questions that are inherently interesting, but will also help to elucidate the function of related genes in other organisms, including humans.

利用从果蝇基因组计划和塞雷拉/应用生物系统公司收集的序列信息，果蝇DNA修复计划已经确定了酵母Rad 51基因的几个旁系同源物。 这些旁系同源物被认为在细胞中DNA损伤的同源重组修复（HRR）过程中促进Rad 51依赖性链交换。 对仓鼠细胞突变的分析，加上鸡细胞中的靶向敲除实验，表明这些基因在提供对电离辐射和化学诱变剂的抗性方面发挥重要作用。 该研究项目的重点是研究DmRad 51 D和DmXRCC 2及其在果蝇有丝分裂同源重组修复中的作用。 长期目标包括使用遗传筛选来鉴定在HRR过程中与DmRad 51 D和DmXRCC 2相互作用的其他基因。 本研究的目的包括：1）证实这些推测的D。通过测定果蝇培养细胞中的功能敲除，2）检查这些基因之间的蛋白质-蛋白质相互作用，3）检查这些基因在果蝇中的表达模式，和4）使用P元件切除以在DmXRCC 2基因中产生功能缺失突变，来鉴定黑腹果蝇Rad 51旁系同源物。DNA包含允许细胞正确运作的指令。 然而，DNA不断地被细胞中的化学物质和紫外线或伽马射线等辐射破坏。 DNA受损的细胞并不总是正常工作，可能导致畸变。 一大群基因修复受损的DNA。 该项目的重点是两个相关基因在DNA修复中的作用。 虽然科学家们对这些基因在试管或单细胞中的作用有了一些了解，但他们仍然不太了解它们在整个生物体中的作用。 该项目将使用分子和遗传工具来研究这些DNA修复基因在果蝇中的作用，其中这些基因的活性可以在整个动物中进行研究。 果蝇在生长和发育过程中使用的几乎所有基因在所有动物中都起着重要作用。 该项目不仅将利用果蝇来探索本质上有趣的问题，而且还将有助于阐明其他生物（包括人类）中相关基因的功能。