Isolation of a coenzyme M-auxotrophic mutant and transformation by electroporation in Methanococcus voltae.
An auxotrophic mutant of Methanococcus voltae was isolated that required coenzyme M (CoM) for growth. With the mutant as a recipient, conditions were developed that allowed the introduction of wild-type CoM+ DNA into the mutant methanogen via electroporation. This method also allowed the rescue of both a histidine and purine auxotroph as well as the introduction of DNA determining resistance to the CoM analog 2-bromoethanesulfonic acid. Electroporation of the CoM(+)-determining DNA was 50- to 80-fold more efficient than natural transformation.
Micheletti
P A
PA
Department of Microbiology, University of Illinois, Urbana 61801.
Sment
K A
KA
Konisky
J
J
D019605:Euryarchaeota / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D015080:Mesna / Q000378:metabolism*; D014170:Transformation, Genetic
D015080:Mesna