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首页> 美国政府科技报告 >Development of Improved Microbial Strains for Methane Production from Municipal Solid Waste and Wastewater
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Development of Improved Microbial Strains for Methane Production from Municipal Solid Waste and Wastewater

机译：改善城市生活垃圾和废水甲烷生产微生物菌株的研究进展

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The overall objective of this sub-program is to apply genetic engineering technology for the development of methanogen-based processes for improved methane production from municipal solid waste (MSW) and waste water. Specifically, the research is directed towards achieving the following goals: (I) the development of a transformation system for Methanobacterium thermoautotrophicum and Methanosarcina mazei; and (II) genetic manipulation of the genes effecting methane generation in M. thermoautotrophicum and Ms. mazei. In the past three years, we have carried out a great deal of work towards achieving the above listed goals. Our major accomplishments during these past three years are: (a) the establishment of a selection method for the development of a transformation system for M. thermoautotrophicum and Ms. mazei; (b) construction of potential cloning vectors for the transformation of M. thermoautotrophicum and Ms. mazei; (c) establishment of ideal conditions for the isolation, purification, and fragmentation of the individual subunits of methylreductase from M. thermoautotrophicum and Ms. mazei. (These will make it possible to analyze part of the amino acid sequence of each subunits to derive the ideal oligonucleotide probes for the isolation of genes encoding these subunits); (d) successful isolation of a number of partial gene fragments via the construction of lacZ fusions. (These gene fragments will be used first to study the necessary genetic elements for gene expression in M. thermoautotrophicum. Eventually, they will be used to construct hybrid genes which can be expressed in both E. coli and M. thermoautotrophicum. Our results also demonstrate that quite a few of the genes from M. thermoautotrophicum can be expressed in E. coli); (e) development of methods for the isolation of promoters from the methanogen genes; and (f) cloning, restriction mapping, and sequence analysis of the cryptic plasmid pME2001 from M. thermoautotrophicum. (ERA citation 13:035908)

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