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, b...

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Bibliographic Details
Published in:Applied microbiology and biotechnology 1994-02, Vol.40 (6), p.864-866
Main Authors: NESVERA, J, HOCHMANNOVA, J, PATEK, M, SROGLOVA, A, BECVAROVA, V
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Brevibacterium - genetics
Electroporation
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Genetic Techniques
Genetic Vectors
Methods. Procedures. Technologies
Modification of gene expression level
Plasmids - genetics
Transformation, Genetic
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Summary: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.
ISSN:0175-7598
1432-0614
DOI:10.1007/bf00173989