A genetic system for Archaea of the genus Methanosarcina : Liposome-mediated transformation and construction of shuttle vectors
- 18 March 1997
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 94 (6) , 2626-2631
- https://doi.org/10.1073/pnas.94.6.2626
Abstract
New methods that allow, for the first time, genetic analysis in Archaea of the genus Methanosarcina are presented. First, several autonomously replicating plasmid shuttle vectors have been constructed based on the naturally occurring plasmid pC2A from Methanosarcina acetivorans. These vectors replicate in 9 of 11 Methanosarcina strains tested and in Escherichia coli. Second, a highly efficient transformation system based upon introduction of DNA by liposomes has been developed. This method allows transformation frequencies of as high as 2 × 108 transformants per microgram of DNA per 109 cells or ≈20% of the recipient population. During the course of this work, the complete 5467-bp DNA sequence of pC2A was determined. The implications of these findings for the future of methanoarchaeal research are also discussed.Keywords
This publication has 40 references indexed in Scilit:
- Plate colonization ofMethanococcus maripaludisandMethanosarcina thermophilain a modified canning jarFEMS Microbiology Letters, 1996
- Pathways of energy conservation in methanogenic archaeaArchiv für Mikrobiologie, 1996
- Transformation ofMethanococcus maripaludisand identification of aPstI-like restriction systemFEMS Microbiology Letters, 1994
- Use of the rep technique for allele replacement to construct new Escherichia coli hosts for maintenance of R6Kλ origin plasmids at different copy numbersGene, 1994
- UNUSUAL COENZYMES OF METHANOGENESISAnnual Review of Biochemistry, 1990
- Construction of an integration vector for use in the archaebacterium Methanococcus voltae and expression of a eubacterial resistance geneMolecular Genetics and Genomics, 1990
- The pMTL nic− cloning vectors. I. Improved pUC polylinker regions to facilitate the use of sonicated DNA for nucleotide sequencingGene, 1988
- Novel regulatory mutants of the phosphate regulon in Escherichia coli K-12Journal of Molecular Biology, 1986
- Mutation to pseudomonic acid resistance ofMethanobacterium thermoautotrophicumleads to an altered isoleucyl-tRNA synthetaseFEMS Microbiology Letters, 1986
- An ancient divergence among the bacteriaJournal of Molecular Evolution, 1977