Genome analysis in Methanosarcina mazei GÖ1: Mechanisms for the adaptation to different carbon- and energy sources

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The mesophilic organism Methanosarcina mazei belongs to the kingdom Archaea and is able to convert H2 + CO2, methanol, methylamines and acetate to methane. Depending on the nature of the carbon- and energy source the organism uses the CO2-reducing, the methylotrophic, or the aceticlastic pathway of methanogenesis. Based on the genomic sequence we would like to analyse which genes are active in a specific pathway, how these genes are regulated and which regulatory networks may be involved. To understand the dynamic nature of gene function the global expression pattern in response to environmental changes will be elucidated. For this purpose genome wide expression profiling will be performed by the use of high-density DNA standard cDNA synthesis from mRNA preparations obtained from cells grown on different substrates or from cells harvested at different growth phases. In the second part of the project defined mutants of Ms. mazei will be created by gene replacement to verify the function of distinct genes. In addition, direct enzymatic assays using cell lysates of Ms. mazei shall contribute to an understanding of cellular processes that directly and indirectly affect methanogenesis.