The regulatory network of nitrogen assimilation and fixation in the methanogenic Archaeon Methanosarcina mazei strain Gö1

产甲烷古菌 Methanosarcina mazei Gö1 菌株氮同化和固定的调控网络

• 批准号: 5314456
• 负责人: Professorin Dr. Ruth Anne Schmitz-Streit
• 金额: --
• 依托单位: Institut für Allgemeine Mikrobiologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2001
• 资助国家: 德国
• 起止时间: 2000-12-31 至 2007-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5314456?language=en
• 关键词: regulatory; network; nitrogen; assimilation; fixation

The mesophilic methanogenic Archaeon Methanosarcina mazei strain Gö1 is able to fix molecular nitrogen when growing under nitrogen-limited conditions. Analyzing the sequence of the genome we have identified and characterized the genes encoding the molybdenum-containing nitrogenase. With this project we seek to analyze the regulatory network of nitrogen metabolism in Ms. mazei using classical genetic approaches and modern molecular tools arising from the knowledge of the complete genome sequence. The first major goal is to identify and to characterize genes, which are regulated by ammonium, using 2-dimensional gel electrophoresis and northern blot analyses. The second goal is to analyze regulatory elements of the nitrogen regulon using two strategies: (i) comparative promoter analyses of the identified nitrogen regulated genes and (ii) identification of promoter binding proteins from cells grown at different nitrogen availabilities by affinity chromatography and gel retardation assays using the identified nitrogen regulated promoter regions. To obtain additional information about the nitrogen regulon and potential cross talks between the nitrogen and the carbon regulon, expression patterns of the whole genome at different nitrogen availabilities and in the presence of different carbon sources will be analyzed by employing DNA-microarrays.

嗜温产甲烷八叠球菌菌株Gö1在氮限制条件下生长时能够固定分子氮。通过对基因组序列的分析，我们鉴定并鉴定了编码含氮固氮酶的基因。在这个项目中，我们试图使用经典的遗传方法和现代分子工具来分析Mazei女士氮代谢的调控网络，这些工具来自于完整基因组序列的知识。第一个主要目标是使用二维凝胶电泳和北方印迹分析来鉴定和表征受铵调节的基因。第二个目标是使用两种策略来分析氮调节子的调节元件：（i）对所鉴定的氮调节基因进行比较启动子分析和（ii）使用所鉴定的氮调节启动子区域通过亲和色谱和凝胶阻滞测定来鉴定来自在不同氮可用性下生长的细胞的启动子结合蛋白。为了获得关于氮调节子和氮与碳调节子之间的潜在串扰的额外信息，将通过使用DNA微阵列分析在不同氮可用性和在不同碳源存在下的全基因组的表达模式。