Open AccessTransformation of Methanococcus maripaludis and identification of a Pst I‐like restriction system
Author(s) -
Tumbula Debra L.,
Makula Ronald A.,
Whitman William B.
Publication year - 1994
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1994.tb07118.x
Subject(s) - psti , methanococcus , plasmid , transformation (genetics) , microbiology and biotechnology , restriction enzyme , biology , genetics , chemistry , dna , gene , archaea
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 3×10 9 cells was 4.8×10 −5 transformants cfu −1 , or 1.8×10 5 transformants μg −1 , which was four orders of magnitude greater than with the natural transformation method. A Pst I restriction activity in M. maripaludis was also identified. Methylation of the plasmid with Pst I methylase increased the methanococcal transformation frequency at least four‐fold. Also, chromosomal DNA from M. maripaludis was resistant to digestion by the Pst I endonuclease.