title: Transfer of the broad-host-range IncQ plasmid RSF1010 and other plasmid vectors to the gram-positive methylotroph Brevibacterium methylicum by electrotransformation.
abstract: Gram-positive facultative methylotrophic coryneform bacterium Brevibacterium methylicum was efficiently transformed with various plasmids using electroporation of intact cells. In addition to the plasmid vectors pEC71 and pZ6-1 constructed on the basis of cryptic plasmids from coryneform bacteria, broad-host-range plasmids pLS5 (derivative of plasmid pMV158 from Streptococcus agalactiae) and RSF1010 belonging to the incompatibility group IncQ from Gram-negative bacteria were found to be present as autonomous structurally unchanged DNA molecules in B. methylicum transformants. With the exception of pZ6-1, all these plasmids were stably maintained in B. methylicum cells grown under non-selective conditions. When plasmid DNAs isolated from B. methylicum were used, the highest efficiency of transformation (10(5) transformants/micrograms DNA) was achieved.
authors.affiliation: Institute of Microbiology, Czech Academy of Sciences, Praha.
authors.forename: J
authors.initials: J
authors.lastname: Nesvera
authors.forename: J
authors.initials: J
authors.lastname: Hochmannová
authors.forename: M
authors.initials: M
authors.lastname: Pátek
authors.forename: A
authors.initials: A
authors.lastname: Sroglová
authors.forename: V
authors.initials: V
authors.lastname: Becvárová
mesh_terms: D001709:Biotechnology; D001951:Brevibacterium / Q000235:genetics*; D018274:Electroporation; D005821:Genetic Techniques; D005822:Genetic Vectors; D010957:Plasmids / Q000235:genetics*; D014170:Transformation, Genetic