Plasmid transformation of Ruminococcus albus by means of high-voltage electroporation.
To apply recombinant DNA techniques to the genetic manipulation of cellulolytic ruminal bacteria, a plasmid vector transformation system must be available. The objective of this work was to develop a system for plasmid transformation of Ruminococcus albus. Using high voltage electrotransformation, pSC22 and pCK17 plasmid vectors, derived from lactic acid bacteria plasmids and replicating via single-stranded DNA intermediate, were successfully introduced into three freshly isolated R. albus strains and into R. albus type strain ATCC 27210. The optimization of the electrotransformation condition raised the electroporation efficiency up to 3 x 10(5) transformants per microgram of pSC22 plasmid.
Cocconcelli
P S
PS
Istituto di Microbiologia, Universitá Cattolica del Sacro Cuore, Piacenza, Italy.
Ferrari
E
E
Rossi
F
F
Bottazzi
V
V
D004269:DNA, Bacterial / Q000235:genetics*; D005822:Genetic Vectors; D006095:Gram-Positive Cocci / Q000235:genetics*; D010957:Plasmids; D014162:Transfection; D014169:Transformation, Bacterial
D004269:DNA, Bacterial