Genetic Analysis of Methanogenesis by Methanosarcina species

拟议研究的长期目标是全面了解甲烷八叠球菌的生理、新陈代谢和进化适应。这项提议的主要重点是确定和描述直接和间接影响使用一碳化合物作为生长和甲烷生产底物的基因。这项研究将(1)研究甲醇是如何调节基因的，(2)研究哪些基因是受甲醇调节的，以及(3)开发初步数据来支持存在一种新的机制来调节甲烷八叠胞菌中蛋白质的产生。拟议的实验将主要利用最近开发的方法进行甲烷八叠球菌的遗传分析；然而，生理、生化和分子方法将补充遗传方法。这项拟议研究的学术价值是基于产甲烷微生物在一些与人类有关的问题上的核心作用，包括从生物材料生产替代燃料、农业、废物处理和全球变暖。拟议的研究有望揭示甲基化营养途径的新方面，通过洞察甲烷生成与有机体的全球新陈代谢的整合。此外，对于古生菌转录和翻译控制的更清楚的理解应该源于对高度调控的甲基化营养基因的检查。拟议研究的更广泛影响包括培训未来的科学家和学者。两名学生将直接得到所需资金的支持。这项研究可能会产生新的遗传工具和方法，这些工具和方法将对更广泛的微生物研究界有用并免费提供。最后，来自其他大学的学生可能会接受本研究期间开发的方法的培训。还将向希望让学生接触微生物世界的当地中小学教师提供设施、资源和建议。

The long-term goal of the proposed research is to understand fully the physiology, metabolism and evolutionary adaptation of Methanosarcina species. The primary emphasis of this proposal is the identification and characterization of genes that directly and indirectly affect the use of one-carbon compounds as substrate for growth and methane production. The research will (1) examine how genes are regulated by methanol, (2) examine which genes are regulated by methanol, and (3) develop preliminary data to support the existence of a novel mechanism to regulate production of proteins in Methanosarcina. The proposed experiments will primarily utilize recently developed methods for genetic analysis of Methanosarcina; however, physiological, biochemical and molecular approaches will complement the genetic approach. The intellectual merit of the proposed research is based on the central role of methanogenic microorganisms in a number of human-related problems including the production of alternative fuels from biological materials, agriculture, waste treatment, and global warming. The proposed studies are expected to reveal novel aspects of the methylotrophic pathway by lending insight into the integration of methanogenesis into the global metabolism of the organism. Moreover, a clearer understanding of archaeal transcriptional and translational control should result from examination of the highly regulated methylotrophic genes. The broader impacts of the proposed research include the training of future scientists and academics. Two students will be directly supported by the requested funds. This study is likely to produce new genetic tools and methods that will be useful, and freely available, to the wider microbial research community. Finally, students from other universities may be trained in the methods developed during this research. Facilities, resources and advice will also be provided to local elementary and secondary school teachers who wish to expose their students to the excitement of the microbial world.