Transfer of the broad-host-range IncQ plasmid RSF1010 and other plasmid vectors to the gram-positive methylotroph Brevibacterium methylicum by electrotransformation.
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.
Nesvera
J
J
Institute of Microbiology, Czech Academy of Sciences, Praha.
Hochmannová
J
J
Pátek
M
M
Sroglová
A
A
Becvárová
V
V
D001709:Biotechnology; D001951:Brevibacterium / Q000235:genetics*; D018274:Electroporation; D005821:Genetic Techniques; D005822:Genetic Vectors; D010957:Plasmids / Q000235:genetics*; D014170:Transformation, Genetic