Transformation of Methanococcus maripaludis and identification of a Pst I-like restriction system

Debra L. Tumbula, Ronald A. Makula, William B. Whitman

Research output: Contribution to journalArticle

92 Scopus citations

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 languageEnglish (US)
Pages (from-to)309-314
Number of pages6
JournalFEMS Microbiology Letters
Volume121
Issue number3
DOIs
StatePublished - Sep 1 1994
Externally publishedYes

Keywords

  • Methanococcus maripaludis
  • Mrr
  • Polyethylene glycol
  • PstI
  • Restriction endonuclease
  • Transformation

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Genetics

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