IRON ASSIMILATION BY PATHOGENIC NEISSERIA SPP

淋病奈瑟菌和脑膜炎奈瑟菌的定植部位 在人类宿主中是多样化的，每个站点代表着一个独特的利基 关于营养、环境因素和竞争 微生物。成长环境对经济增长有显著影响。 淋球菌和脑膜炎奈瑟菌的代谢和细胞组成， 细胞成分的改变通常反映在相互作用中 这些微生物与人类宿主的关系。 这个项目的总体目标是阐明具体的机制。 被致病奈瑟氏菌用于同化精华 营养铁。我们将把大部分努力集中在主要的铁- 调节蛋白，FEP。最近克隆的FEP将允许我们创建 经mini T13诱变产生FEP缺陷的突变株。我们会 检查FEP结构基因的DNA序列5‘，并研究如何 铁调节FEP的表达。我们还将审查这项规定 FEP成绩单的铁杆。 淋球菌FEP在细胞中的具体位置将被确定 通过渗透休克技术，以及它与其他细胞的相互作用 通过交联化技术来合成组份。最后，我们将使用我们最近的 描述了脑膜炎奈瑟氏菌Tn916诱变系统以分离出特定的 铁利用有缺陷的突变体。由此产生的突变DNA将是 用作探针来克隆相应的野生型DNA。 在这些研究中产生的突变将在我们的动物体内进行测试 模特们。将采用小鼠皮下腔模型进行研究。 淋病奈瑟菌和脑膜炎奈瑟氏菌突变体的生长。此外, 我们将在小鼠菌血症模型中检查脑膜炎奈瑟氏菌的突变体。 这些研究将使我们能够评估铁的利用在 淋病奈瑟菌感染后的生长和致病性 和脑膜炎奈瑟氏菌。

The sites colonized by Neisseria gonorrhoeae and Neisseria meningitidis within the human host are diverse and each site represents a unique niche with respect to nutrients, environmental factors and competing microorganisms. The growth environment has a marked effect on the metabolism and cellular composition of N. gonorrhoeae and N. meningitidis, and an altered cellular composition is often reflected in the interaction of these microorganisms with the human host. The overall goal of this project is to elucidate the specific mechanisms employed by the pathogenic Neisseria for the assimilation of the essential nutrient iron. We will focus a majority of our efforts on the major iron- regulated protein, Fep. The recently cloned fep will allow us to create mutants defective in the production of Fep by miniT13 mutagenesis. We will examine the DNA sequences 5' to the fep structural gene and investigate how iron regulates the expression of Fep. We will also examine the regulation by iron of the Fep transcript. The specific location of the gonococcal Fep in the cell will be determined by osmotic shock techniques, as well as its interaction with other cellular components by cross-linking techniques. Finally, we will use our recently described N. meningitidis Tn916 mutagenesis system to isolate specific mutants defective in iron utilization. The resulting mutated DNA will be used as probes to clone the corresponding wildtype DNA. Mutants generated in these studies will be examined in vivo in our animal models. The mouse subcutaneous chamber model will be employed for studies on the growth of N. gonorrhoeae and N. meningitidis mutants. In addition, we will examine N. meningitidis mutants in the mouse bacteremia model. These studies will allow us to evaluate the role of iron utilization in the growth and subsequent pathogenicity of infection caused by N. gonorrhoeae and N. meningitidis.