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：这项提案涉及基因的遗传分析。 原核细胞中细胞分裂所必需的。的最终目标是 该项目是为了完全了解这种复合体的分子机制 基本流程。特别是，细胞质分裂是在 丝状土壤细菌--天蓝色链霉菌 服从基因分析的。天南星是一种特别有利的 用于识别参与细菌胞质分裂的基因的系统，因为 细胞分裂，通常是一个必要的过程，对于生长和 这种菌丝体的生存能力。这样做的目的是隔离关键的 尚未在任何原核生物中发现的基因。这个 该项目确定的基因产物可能成为潜在的目标 具有生物活性的新型化合物。对这种化合物的需求是 在出现多重耐药的今天尤其重要 病原体。 该项目建议识别和表征与细胞有关的基因。 采用两种诱变和克隆策略的部门。在第一个 方法，分裂基因将通过化学诱变和 利用有序粘粒互补文库克隆。在第二个 方法，将通过插入的方式鉴定和克隆分裂基因 利用转座子进行诱变。将使用免疫荧光显微镜 为了确定关键分裂蛋白FtsZ的定位是否 分布在新的突变体中。影响这个早期事件的基因是 特别感兴趣，因为它们还没有被任何生物体识别出来。 最后，提出了实验来分析新的产品是否 已识别的基因共定位在相同的亚细胞位置和模式 FtsZ。