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.
Original language | English (US) |
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Pages (from-to) | 309-314 |
Number of pages | 6 |
Journal | FEMS Microbiology Letters |
Volume | 121 |
Issue number | 3 |
DOIs | |
State | Published - Sep 1 1994 |
Externally published | Yes |
Keywords
- Methanococcus maripaludis
- Mrr
- Polyethylene glycol
- PstI
- Restriction endonuclease
- Transformation
ASJC Scopus subject areas
- General Medicine