Genetic Analysis of Methylotrophic Methanogenesis in Methanosarcina Species

甲烷八叠球菌属甲基营养型产甲烷作用的遗传分析

• 批准号: 0212466
• 负责人: William Metcalf
• 金额: $ 36万
• 依托单位: University of Illinois at Urbana-Champaign
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2005-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0212466&HistoricalAwards=false
• 关键词: Genetic; Analysis; Methylotrophic; Methanogenesis; Methanosarcina

The long-term goal of the research is to expand our knowledge regarding the physiology, metabolism and evolutionary adaptation of the methane-producing Methanosarcina species. Although the primary emphasis of these studies will be the identification and characterization of genes that directly and indirectly effect the utilization of methanol as a growth substrate (methylotrophic methanogenesis), the results are expected to have broad implications with respect to Methanosarcina and other methanogenic organisms. In particular, it is expected that these studies will reveal not only novel aspects of the methylotrophic pathway, but also how methanogenesis is integrated into the overall metabolism of the organism. Further, it is expected that a clearer understanding of archaeal transcriptional control will result from examination of the highly regulated methylotrophic genes. The experiments will be primarily based on recently developed methods for genetic analysis of Methanosarcina species; however, physiological, biochemical and molecular approaches will also be utilized to complement the genetic studies. This approach is expected to generate data that are both novel and complementary to the results of preceding studies. To achieve these aims Methanosarcina strains with mutations in a number of genes believed to be involved in methylotrophic methanogenesis will be constructed. These mutant strains will then be characterized with respect to a variety of physiological and biochemical traits. Random mutagenesis and proteomics approaches will be used to identify new genes that are specifically required for growth on methylotrophic substrates. Separate studies will examine the regulation of genes known to be involved and attempt to identify the genes that mediate this regulation. Finally, during the course of this work additional methods will be developed for genetic analysis of Methanosarcina species.

这项研究的长期目标是扩大我们对产甲烷八叠球菌的生理、代谢和进化适应的认识。虽然这些研究的主要重点将是识别和表征的基因，直接和间接影响利用甲醇作为生长底物（甲基营养型甲烷），结果预计将有广泛的影响甲烷八叠球菌和其他产甲烷生物。特别是，预计这些研究不仅将揭示甲基营养途径的新方面，而且还将揭示甲烷生成如何整合到生物体的整体代谢中。此外，它是预期的古细菌转录控制的更清楚的理解将导致高度调节的甲基营养基因的检查。实验将主要基于最近开发的甲烷八叠球菌属物种遗传分析方法;然而，生理学、生物化学和分子方法也将用于补充遗传研究。这种方法有望产生新的数据，并对先前研究的结果进行补充。为了实现这些目标，将构建在许多被认为参与甲基营养型甲烷生成的基因中具有突变的甲烷八叠球菌菌株。这些突变株将被表征为具有多种生理和生化特性。随机诱变和蛋白质组学方法将被用来确定新的基因，特别是甲基营养基质上的生长所需的。单独的研究将检查已知参与的基因的调节，并试图确定介导这种调节的基因。最后，在这项工作的过程中，其他方法将开发甲烷八叠球菌属物种的遗传分析。