Transformation of Methanococcus maripaludis and identification of a Pst I-like restriction system
- 1 September 1994
- journal article
- Published by Oxford University Press (OUP) in FEMS Microbiology Letters
- Vol. 121 (3) , 309-314
- https://doi.org/10.1016/0378-1097(94)90309-3
Abstract
An optimized polyethylene glycol (PEG) method of transformation was developed for Methanococcus maripaludis using the pKAS102 integration vector. The frequency of transformation with 0.8 μg of plasmid and 3×109 cells was 4.8×10−5 transformants cfu−1, or 1.8×105 transformants μg−1, which was four orders of magnitude greater than with the natural transformation method. A PstI restriction activity in M. maripaludis was also identified. Methylation of the plasmid with PstI methylase increased the methanococcal transformation frequency at least four-fold. Also, chromosomal DNA from M. maripaludis was resistant to digestion by the PstI endonuclease.Keywords
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