Applied Microbiology and Biotechnology, Vol.40, No.6, 864-866, 1994
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/mu g DNA) was achieved.
Keywords:CORYNEBACTERIUM-GLUTAMICUM;MYCOBACTERIUM-SMEGMATIS;ESCHERICHIA-COLI;TRANSFORMATION;ELECTROPORATION;GENES;DNA