DNA Replication in B. subtilis

枯草芽孢杆菌中的 DNA 复制

• 批准号: 6773903
• 负责人: CHARLES S MCHENRY
• 金额: $ 29.93万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6773903
• 关键词: Bacillus subtilis; DNA binding protein; DNA replication; DNA replication origin; bacterial RNA; bacterial genetics; bacterial proteins; gram positive bacteria; helicase; immunoaffinity chromatography; laboratory rabbit; molecular cloning; polymerase chain reaction; southern blotting; structural genes; western blottings

DESCRIPTION (provided by applicant): Decades of study have provided us with detailed information about DNA replication in E.coli. The explosion of bacterial genomic sequences has enabled facile exploration beyond the standard E. coil prototype. The objective of these proposed studies is to establish a complete replication system for a second very distant bacterium, B. subtilis. B. subtilis is ideally suited for these purposes because i. it is diverse, belonging to the low GC Gram (+) classification, evolutionarily distant (>>1 billion yr.) from Proteobacteria and E. coli, ii. its genome has been sequenced and it has a developed genetic system, iii. it can be grown in fermentors in large quantity making it suitable for biochemical analysis, iv) genomics indicates it is representative of the most diverse group of bacteria from a DNA replication standpoint (having two apparent distinct replicases and a number of replication proteins with no counterpart in E. coli, and v) it provides a model system representative of an important group of important human pathogens (streptococci, staphylococci and enterococci. The studies for the proposed grant period build upon a strong base established through unfunded preliminary studies and will focus upon establishing i. the identity of the elongation apparatus components and function, ii. the proteins required for providing RNA primers and iii. the proteins required for origin-specific initiation. Emphasis will be placed on those features of the B. subtilis replication system that make it distinct from E. coli. This will provide important information regarding the variations that can be expected between bacterial replication systems in terms of basic components, mechanisms and regulation. Understanding the biochemical details of DNA replication in an organism that contains two DNA polymerase Ills, may provide paradigms applicable to other two-polymerase replicases including eukaryotes where Pol epsilon and delta both appear to be involved in processive chromosomal replication.

描述（由申请人提供）： 数十年的研究为我们提供了有关E.Coli中DNA复制的详细信息。细菌基因组序列的爆炸使超出标准的E.线圈原型超出了便利探索。这些提出的研究的目的是建立第二个非常遥远的细菌的完整复制系统，即枯草芽孢杆菌。 B.枯萎病非常适合这些目的，因为i。它是多种多样的，属于低GC革兰（+）分类，从蛋白质细菌和大肠杆菌，II。它的基因组已被测序，并具有开发的遗传系统III。它可以大量生长在发酵罐中，使其适合生化分析，iv）基因组学表明，它是从DNA复制的角度来代表最多样化的细菌群体（具有两个明显不同的复制酶，并且在E. coli和V中没有对应物，并且在E. coli和V中没有任何对应的人类代表（IT）的重要人类代表（IT）的重要人类（IT），这是重要的人类代表，重要的是重要的人类。葡萄球菌和肠球菌的授予时期的研究是建立不受欢迎的初步研究的，并将重点放在建立延长设备的身份上。大肠杆菌将提供有关细菌复制系统在基本组件，机制和调节方面可以预期的变化的重要信息。了解包含两个DNA聚合酶疾病的生物体中DNA复制的生化细节，可以提供适用于其他两种聚合酶复制酶在内的范式，包括真核生物，其中Pol Epsilon和Delta似乎都参与了过程中的染色体染色体复制。