Transformation of a Methylotrophic Bacterium, Methylobacterium extorquens, with a Broad-Host-Range Plasmid by Electroporation

Appl Environ Microbiol. 1991 Apr;57(4):924-6. doi: 10.1128/aem.57.4.924-926.1991.

Abstract

An efficient method for the transformation of the methylotrophic bacterium Methylobacterium extorquens NR-2 with a broad-host-range plasmid, pLA2917, by electroporation was examined. Transformants of M. extorquens NR-2 expressing resistance to kanamycin were obtained after electric pulse. These transformants were found to harbor a single plasmid which was electrophoretically identical and homologous to pLA2917 obtained from Escherichia coli. Several factors which determined the transformation efficiency were optimized, resulting in a transformation efficiency of up to 8 x 10 transformants per mug of plasmid DNA by 10 pulses at a field strength of 10 kV/cm and a pulse duration of 300 mus.