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：该提案涉及基因的遗传分析 原核细胞的细胞分裂所需。 该项目的最终目标是 项目的目的是彻底了解这种复合物的分子机制 基本过程。 特别是，胞质分裂的研究 丝状土壤细菌 天蓝色链霉菌 (Streptomyces coelicolor)，一种容易 适合遗传分析。 天蓝色链球菌 (S. coelicolor) 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重要的过程，对于生长和发育来说是可有可无的 该菌丝体的活力。 目的是隔离关键的 尚未在任何原核生物中鉴定出的基因。 这 该项目确定的基因产物可能是潜在的目标 具有生物活性的新型化合物。 对此类化合物的需求是 随着多重耐药性的出现，这一点在今天尤为重要 病原体。 该项目旨在鉴定和表征细胞中涉及的基因 采用两种诱变和克隆策略进行分裂。 在第一个 方法，分裂基因将通过化学诱变和 使用有序粘粒互补文库进行克隆。 在第二个 方法，将通过插入来鉴定和克隆分裂基因 使用转座子进行诱变。 将使用免疫荧光显微镜 确定关键分裂蛋白 FtsZ 的定位是否 分布在新的突变体中。 影响这一早期事件的基因是 特别感兴趣，因为它们尚未被任何生物体识别。 最后，提出实验来分析新产品是否 鉴定出的基因共定位于相同的亚细胞位置和模式 FtsZ。