Methanosarcina Genomics

该项目将利用最近完成的醋酸甲烷八叠球菌基因组序列来识别两个关键生理过程的新基因/蛋白质。 本研究将采用功能基因组学的方法来进一步了解（i）M. acetivorans，和（ii）M.食醋动物和食醋动物。测序的微生物基因组的大部分开放阅读框与任何已知蛋白质没有显著的推导同一性;因此，预计将发现新的蛋白质和酶，并且它们的表征将揭示新的生物化学原理。将采用全基因组DNA微阵列和二维凝胶电泳方法来鉴定在乙酸盐与甲醇或甲胺生长期间以及在含有高盐和低盐的培养基中差异表达的基因。预计结果将（i）鉴定出醋酸盐形成甲烷的特异性基因;（ii）揭示古细菌适应不断变化的环境条件的一般原则。将通过转录图谱对差异表达的基因进行表征，并分析基因序列以获得有关其潜在功能的信息。将进行基因敲除实验，以验证其在乙酸发酵途径或渗透胁迫中的作用，并帮助确定其特定功能。选择的基因将被过度表达，基因产物将被表征以确定它们的生化特性和特定的生理功能。通过更详细地阐明生理过程，该项目有望通过揭示M.乙酰基食腐菌对不断变化的环境条件以及它如何与构成全球碳循环中厌氧环节的微生物聚生体中的其他成员相互作用作出反应。 此外，来自每个学术机构的学生将接触到来自每个合作学术机构的各种实验方法和方法。研究的基本互动性质将为参与的学生提供广泛的接触遗传学，基因组学，发酵，微生物生理学和酶表征。这种类型的广泛培训旨在为未来的研究提供人员，这些研究将基于未来十年及以后预期的大量基因组信息

This project will exploit the recently completed genomic sequence of Methanosarcina acetivorans to identify novel genes/proteins for two key physiological processes. A functional genomics approach will be taken to further the understanding of (i) the fermentation of acetate to methane (the pathway by which two-thirds of the global output of methane is produced) by M. acetivorans, and (ii) the response to environmental stress by M. acetivorans and the Archaea in general. A large percentage of the open reading frames of sequenced microbial genomes have no significant deduced identity to any known proteins; thus it is expected that novel proteins and enzymes will be discovered and that their characterization will uncover new biochemical principles. Whole genome DNA microarray and two-dimensional gel electrophoresis methods will be employed to identify genes that are differentially expressed during growth on acetate versus methanol or methylamines, and in media containing high and low salt. The results are expected to (i) identify genes specific for methane formation from acetate and (ii) uncover general principles of Archaeal adaptation to changing environmental conditions. The differentially expressed genes will be characterized by transcriptional mapping and the gene sequences will be analyzed for information regarding their potential function. Gene knockout experiments will be performed to validate their role in the acetate fermentation pathway or osmotic stress, and also to aid in the determination of their specific function. Selected genes will be over-expressed and the gene products characterized to determine their biochemical properties and specific physiological functionBy elucidating physiological processes in greater detail, the project is expected to increase the understanding of biocomplexity by revealing how M. acetivorans reacts to changing environmental conditions and how it interacts with other members in microbial consortia that comprise the anaerobic link in the global carbon cycle. In addition, students from each academic institution will be exposed to a wide variety of experimental approaches and methods from each of the collaborating academic institutions . The fundamental interactive nature of the research will provide participating students with broad exposure to genetics, genomics, fermentation, microbial physiology, and enzyme characterization. This type of broad training is designed to provide personnel for future research that will be based on the plethora of genomic information expected in the next decade and beyond