GENETIC DISSECTION OF BACTERIAL CYTOKINESIS

细菌细胞分裂的基因解剖

• 批准号: 6386806
• 负责人: JOSEPH R MC CORMICK
• 金额: $ 9.7万
• 依托单位: DUQUESNE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6386806
• 关键词: Streptomyces; bacterial genetics; cell cycle; cell motility; gene mutation; genetic library; immunofluorescence technique; molecular cloning; protein localization; site directed mutagenesis; transposon /insertion element

DESCRIPTION: This proposal is concerned with the genetic analysis of genes required for cell division in prokaryotic cells. The ultimate goal of the project is to completely understand the molecular mechanism of this complex fundamental process. In particular, cytokinesis is studied in the filamentous soil bacterium Streptomyces coelicolor, a species readily amenable to genetic analysis. S. coelicolor is a particularly advantageous system to use to identify genes involved in bacterial cytokinesis because cell division, normally an essential process, is dispensable for growth and viability in this mycelial organism. The intent is to isolate critical genes that have not yet been identified in any prokaryotic organism. The products of genes identified by this project could be potential targets for novel compounds with biological activity. The need for such compounds is especially important today with the emergence of multiply drug-resistant pathogens. This project proposes to identify and characterize genes involved in cell division employing two mutagenesis and cloning strategies. In the first approach, division genes will be identified by chemical mutagenesis and cloned using ordered cosmid complementation libraries. In the second approach, division genes will be identified and cloned by insertional mutagenesis using a transposon. Immunofluorescence microscopy will be used to determine if the localization of FtsZ, a key division protein, is distributed in the new mutants. Genes that affect this early event are of particular interest because they have not been identified by any organism. Finally, experiments are proposed to analyze if the products of newly identified genes co-localize at the same subcellular position and pattern as FtsZ.

描述：本提案涉及基因的遗传分析 是原核细胞分裂所必需的。 的最终目标 该项目的目的是完全了解这种复合物的分子机制 基本过程。 特别地，胞质分裂是在 丝状土壤细菌天蓝色链霉菌，一种容易 适合基因分析。 S.腔棘鱼是一种特别有利的 该系统用于鉴定涉及细菌胞质分裂的基因， 细胞分裂，通常是一个基本过程，是生长的关键， 在这种菌丝体中的生存能力。 目的是隔离关键的 这些基因在任何原核生物中都没有被发现。 的 该项目确定的基因产物可能是潜在的靶点， 具有生物活性的新化合物。 对此类化合物的需求是 尤其重要的是，随着多重抗药性的出现， 病原体 该项目提出识别和表征参与细胞凋亡的基因， 使用两种诱变和克隆策略进行分裂。 上 方法，分裂基因将通过化学诱变鉴定， 使用有序粘粒互补文库克隆。 在第二 方法，分裂基因将被鉴定和克隆的插入 使用转座子的诱变。 将使用免疫荧光显微镜检查 以确定FtsZ（一种关键分裂蛋白）的定位是否 分布在新的突变体中。 影响这一早期事件的基因是 特别值得注意的是，它们还没有被任何生物识别出来。 最后，通过实验分析了新产品的性能 鉴定的基因共定位在相同的亚细胞位置和模式， FtsZ。