Analysis of Oxygen Adaptation in Methanogenic Archaea

产甲烷古菌的氧适应分析

• 批准号: 6791500
• 负责人: DANIEL J LESSNER
• 金额: $ 4.3万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6791500
• 关键词: Archaea; anaerobic bacteria; bacterial genetics; free radical oxygen; gene expression; gene expression profiling; genetic regulation; microarray technology; northern blottings; oxidative stress; oxidoreductase; oxygen consumption; polymerase chain reaction; postdoctoral investigator; protein structure function

DESCRIPTION (provided by applicant): The long term objectives of the proposed research are to identify and characterize novel proteins associated with oxygen adaptation in anaerobic methane-producing Archaea in order to provide a foundation of the function and evolution of oxygen detoxification enzymes. This investigation is important due to the relevance of detoxification of oxidative species as a general defense mechanism in all living cells, but also in the response of pathogens to oxidative burst produced by host defense cells. In addition, methane production is relevant to the problems of global warming and there is still much to learn about the physiology, biochemistry, and genetics of Archaea. The specific aims of this research proposal are: a) utilize a Methanosarcina acetivorans complete genome microarray to identify genes up-regulated during exposure of cells to oxygen, superoxide, and hydrogen peroxide, b) conduct transcriptional analysis and expression profiling of a previously identified gene cluster encoding probable oxidative stress proteins, including a superoxide reductase and rubredoxin: oxygen oxidoreductase (ROO), c) overexpress and purify the putative ROO from M. acetivorans to verify its catalytic properties in the context of the physiology of this organism.

描述（由申请人提供）：所提议研究的长期目标是鉴定和表征与厌氧产甲烷古细菌中的氧适应相关的新型蛋白质，以便为氧解毒酶的功能和进化提供基础。这项研究很重要，因为氧化物质的解毒作为所有活细胞中的一般防御机制，而且在病原体对宿主防御细胞产生的氧化爆发的反应中也具有相关性。此外，甲烷的产生与全球变暖问题有关，而且古细菌的生理学、生物化学和遗传学还有很多东西需要了解。该研究计划的具体目标是：a) 利用乙酸甲烷八叠球菌完整基因组微阵列来识别细胞暴露于氧气、超氧化物和过氧化氢期间上调的基因，b) 对先前识别的编码可能的氧化应激蛋白（包括超氧化物还原酶和红氧还蛋白：氧）的基因簇进行转录分析和表达谱分析 氧化还原酶 (ROO)，c) 过表达并纯化来自 M. acetivorans 的假定 ROO，以验证其在该生物体生理学背景下的催化特性。