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.