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OSMOREGULATION IN STAPHYLOCOCCUS AUREUS

金黄色葡萄球菌的渗透压调节

基本信息

批准号：
3438788
负责人：
BRIAN JAMES WILKINSON
金额：
$ 10.15万
依托单位：
ILLINOIS STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1991
资助国家：
美国
起止时间：
1991-05-01 至 1994-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3438788
关键词：
Staphylococcus aureus aminoacid transport deficient growth media ecology gene expression genetic manipulation genetic transcription gram positive bacteria growth media microorganism culture microorganism growth molecular cloning mutant osmotic pressure radiochemistry transposon /insertion element

项目摘要

The broad long-term objective of this proposal is to provide a comprehensive description of osmoregulation in Staphylococcus aureus. It is hoped that this knowledge will have wide application to gram positive bacteria. S. aureus is an extremely halotolerant eubacterium capable of growing in media containing in excess of 3 M NaCl. The physiological mechanisms allowing this remarkable osmotolerance are poorly understood. Also, growth with NaCl is known to profoundly affect the cell physiology of S. aureus. S. aureus is a major cause of food poisoning and an extremely versatile pathogen of a variety of organs and tissues. Ability to grow in high osmolarity environments might be an important attribute of the pathogenicity of the organism. The physiology and biochemistry of osmoregulation will be studied by determining the identity and intracellular concentration of molecules (osmolytes) accumulating during growth in high osmolarity media. The dynamics of osmoregulation will be probed in experiments where cells are subjected to hyperosmotic stress. Transport systems for osmolytes will be studied in terms of their kinetics, substrate specificity and response to osmotic stress in whole cells and membrane vesicle preparations. Genetic studies will involve the creation of various osmolyte mutants by chemical and transposon mutagenesis. Osmolyte transport or metabolism mutants will be selected for inability to grow on defined medium containing 2 M NaCl plus an osmolyte (e.g., proline, glycine betaine, choline). Exogenous osmolyte-independent mutants, and proline analog-resistant transport mutants will also be created. Transposon lacZ fusion mutants will be used to study the transcriptional regulation of osmoregulatory genes. Transposon insertion mutants will be used to clone genes involved in osmoregulation.

这项建议的长远目标是提供一个金黄色葡萄球菌中的细菌生长调节的综合描述。希望这些知识能广泛应用于克阳性细菌S.金黄色葡萄球菌是一种极端耐盐的真细菌能够在含有超过3M NaCl的培养基中生长。的允许这种显著的抗氧化耐受性的生理机制是不太了解。此外，已知NaCl的生长会深刻影响 S.金黄色。 S.金黄色葡萄球菌是食物中毒的主要原因中毒和一个非常多才多艺的病原体的各种器官，组织中在高渗透压环境中生长的能力可能是病原体致病性的重要属性。的将通过以下方式研究调节的生理学和生物化学确定分子的身份和细胞内浓度（渗透压调节剂）在高渗透压培养基中生长期间积累。的将在实验中探索细胞的调节动力学，经受高渗压力。渗透剂的运输系统将在动力学、底物特异性和全细胞和膜囊泡对渗透胁迫的反应准备工作遗传学研究将涉及创造各种通过化学和转座子突变产生渗透压突变体。压剂运输或代谢突变体将被选择为不能生长在含有2M NaCl加渗压剂（例如，脯氨酸，甘氨酸甜菜碱、胆碱）。外源性不依赖渗透压调节剂的突变体，以及还将产生脯氨酸类似物抗性转运突变体。转座子lacZ融合突变体将被用于研究转录调控基因的表达。转座子插入突变体将可用于克隆参与转录调控的基因。