GENETIC DISSECTION OF BACTERIAL CYTOKINESIS

细菌细胞分裂的基因解剖

• 批准号: 6525427
• 负责人: JOSEPH R MC CORMICK
• 金额: $ 9.88万
• 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 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6525427
• 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：该建议与基因的遗传分析有关 核细胞中细胞分裂所必需的。 最终目标 项目是完全了解该复合物的分子机制 基本过程。 特别是，在 丝状土壤细菌链霉菌卵石，一种容易的物种 适合遗传分析。 S. coelicolor是一个特别有利的 用于鉴定细菌细胞因子涉及的基因的系统，因为 细胞分裂通常是一个基本过程，可用于生长和 这种菌丝生物的生存能力。 目的是隔离关键 尚未在任何原核生物中鉴定出来的基因。 这 该项目确定的基因产品可能是潜在的目标 具有生物活性的新型化合物。 对这种化合物的需求是 今天尤其重要，随着抗药性的出现 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 使用有序的宇宙互补库克隆。 在第二个 方法，分裂基因将被插入识别和克隆 使用转座子的诱变。 免疫荧光显微镜将使用 确定ftsz的定位（关键分裂蛋白）是否是 分布在新突变体中。 影响这一早期事件的基因是 特别的兴趣是因为它们尚未被任何生物体识别。 最后，提出了实验来分析新的产品是否新近 鉴定的基因在相同的亚细胞位置和模式下共定位 ftsz。