DNA Replication in Halobacterium sp. NRC-1

盐杆菌属 DNA 复制。

• 批准号: 0450695
• 负责人: Shiladitya DasSarma
• 金额: --
• 依托单位: University of Maryland Biotechnology Institute
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-15 至 2010-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0450695&HistoricalAwards=false
• 关键词: DNA; Replication; Halobacterium; sp.; NRC

Halobacterium sp. NRC-1 is a model system for studies of Archaea and extremophiles. DNA replication is of special interest in this Archaeon because it can serve as a simple model for replication in eukaryotes and because it is relatively poorly understood in Archaea. The replication origins from the chromosome of the model archaeaon, Halobacterium NRC-1, and the common region of the large extrachromosomal replicons, pNRC100 and pNRC200 have been cloned. Additional bioinformatic analysis has identified a set of likely genes involved in DNA replication. The goal of this project is to further examine the cis and trans acting factors that are important for DNA replication initiation in Halobacterium NRC-1. The cis acting regions will be studied by detailed random and site-directed mutagenesis of previously identified autonomously replicating sequence (ARS) elements, in order to establish the essential regions and their functions. The protein factors which bind to the conserved ARS element sequences will be identified by biochemical methods, in vivo and in vitro. A second goal of the project is to establish which genomic regions are used to initiate DNA replication in vivo, their directionality, and the mechanism of coordinate regulation of the three replicons in the Halobacterium NRC-1 genome. For this goal, DNA microarrays and genetic methodologies will be used. A third goal of the project is to address the essentially of genes predicted to be involved in the initiation of DNA replication using a recently improved genetic knockout system. This project will provide valuable insights into a fundamental and relatively poorly studied process in the third branch of life. A deeper appreciation of the diversity of microbes, especially extremophiles, will result, with possible implications for biotechnology. This project will contribute to the development of human resources in an important area of biological research, including the training of graduate students and involvement of other non-scientists through outreach. The ultimate goal is to bring the understanding of the players and processes of DNA replication in an archaeon closer to the level of understanding in bacterial and eukaryotic model systems.

盐杆菌NRC-1是一个研究极端微生物和盐生菌的模式系统。 DNA复制在这一领域特别重要，因为它可以作为真核生物复制的一个简单模型，而且在真核生物中对它的了解相对较少。 已克隆了模式古细菌嗜盐杆菌NRC-1染色体的复制起点和大的染色体外复制子pNRC 100和pNRC 200的共同区域。额外的生物信息学分析已经确定了一组可能参与DNA复制的基因。本项目的目的是进一步研究盐杆菌NRC-1中对DNA复制起始起重要作用的顺式和反式作用因子。顺式作用区域将通过先前鉴定的自主复制序列（ARS）元件的详细随机和定点诱变来研究，以建立必需区域及其功能。将通过生物化学方法在体内和体外鉴定与保守ARS元件序列结合的蛋白因子。该项目的第二个目标是确定哪些基因组区域用于启动体内DNA复制，它们的方向性以及盐杆菌NRC-1基因组中三个复制子的协调调节机制。 为此，将使用DNA微阵列和遗传学方法。 该项目的第三个目标是使用最近改进的基因敲除系统来解决预测参与DNA复制启动的基因的本质。这个项目将提供有价值的见解，一个基本的和相对较少研究的过程中的第三个分支的生活。 对微生物的多样性，特别是极端微生物的多样性的更深入的了解，将可能对生物技术产生影响。 这一项目将有助于在生物学研究的一个重要领域开发人力资源，包括培训研究生和通过外联活动让其他非科学家参与。 最终目标是使古细菌中DNA复制的参与者和过程的理解更接近细菌和真核模型系统的理解水平。