STUDIES OF BACTERIAL MINICELL MUTANTS

这个项目的长期目标是了解这种机制。 细菌细胞用来将分裂部位放置在合适的位置 在中层细胞。为了达到这一目标，细菌微细胞基因座 (MinB)将是研究，因为我们之前已经表明，三分钟 基因产物在正常的细胞周期中起着重要的作用 确保中细胞处的隔膜放置正确。这是 由除法抑制器的联合作用完成，取决于 Minc和Mind蛋白的存在以及拓扑特异性 因子，矿物质蛋白质。遗传学、免疫化学和 免疫电子显微镜和生物化学将用于 以下目标：(1)确定三者的细胞定位 MIN蛋白在正常条件下和在以下条件下 间隔位置的拓扑特异性被扰动；(2) 确定哪些蛋白质与细胞内的Min蛋白相互作用； (3)确定在我的领域内负责其 能够为选址过程提供拓扑特异性， 并使用遗传手段来鉴定起作用的细胞蛋白质 在拓扑专用性函数中；(4)利用遗传手段 鉴定在Minc介导的蛋白质中起作用的细胞蛋白质 隔阻断；(5)测定Minc-ATPase活性。 介导性间隔阻断；(5)确定ATPase活性如何 思维与它在选址过程中的作用是相联系的。

The long-range objective of this project is to understand the mechanism used by bacterial cells to place the division site at its proper location at midcell. To approach this objective, the bacterial minicell locus (minB) will be studies since we have previously shown that the three min gene products play an important role during the normal cell cycle to ensure the proper placement of the division septum at midcell. This is accomplished by the combined action of a division inhibitor, dependent on the presence of the MinC and MinD proteins, and a topological specificity factor, the MinE protein. A combination of genetics, immunochemistry and immunoelectron microscopy, and biochemistry will be used, with the following goals: (1) to determine the cellular localization of the three Min proteins under normal conditions and under conditions where the topological specificity of septal placement is perturbed; (2) to determine which proteins interact with the Min proteins within the cell; (3) to identify the domain within MinE that is responsible for its ability to give topological specificity to the site-selection process, and to use genetic means to identify cellular proteins that play a role in the topological specificity function of MinE; (4) to use genetic means to identify cellular proteins that play a role in the MinC-mediated septation block; (5) to determine how the ATPase activity of MinC- mediated septation block; (5) to determine how the ATPase activity of MinD is coupled to its role in the site-selection process.