Development of a Gene Cloning System in Methanogens
Abstract
The methane-producing archaebacteria have a unique biochemistry and molecular biology. In this work, a collection of mutants in the thermophilic autotroph Methanobacterium thermoautotrophicum was obtained. Several of these were resistant to pyrimidine and purine analogs (e.g. 5-fluorouracil and 6- mercaptopurine). High form wild type cells to resistance. A formate auxotroph was isolated, which represents a novel growth requirement. Characterization of this stain will reveal new aspects of one-carbon metabolism in methanogens. A complete genomic library of restriction fragments of the methanogen DNA was prepared in Escherichia coli plasmid pBR322. Transposon sequences will be introduced into these fragments, and linearized, transposon-containing plasmids used to transform the methanogen. This technique (additive transformation) followed by selective enrichment will yield mutants. The mutants and the ability to introduce DNA into recipient strains will provide the basis for genetic mapping and engineering of methanogens. Keywords: Gene cloning.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 1988
- Accession Number
- ADA197062
Entities
People
- David Mccarthy
- David P. Nagle Jr.
Organizations
- University of Oklahoma