Biolistic transformation of a procaryote, Bacillus megaterium.
We present a simple and rapid method for introducing exogenous DNA into a bacterium, Bacillus megaterium, utilizing the recently developed biolistic process. A suspension of B. megaterium was spread onto the surface of nonselective medium. Plasmid pUB110 DNA, which contains a gene that confers kanamycin resistance, was precipitated onto tungsten particles. Using a biolistic propulsion system, the coated particles were accelerated at high velocities into the B. megaterium recipient cells. Selection was done by use of an agar overlay containing 50 micrograms of kanamycin per ml. Antibiotic-resistant transformants were recovered from the medium interface after 72 h of incubation, and the recipient strain was shown to contain the delivered plasmid by agarose gel electrophoresis of isolated plasmid DNA. All strains of B. megaterium tested were successfully transformed by this method, although transformation efficiency varied among strains. Physical variables of the biolistic process and biological variables associated with the target cells were optimized, yielding greater than 10(4) transformants per treated plate. This is the first report of the biolistic transformation of a procaryote.
Shark
K B
KB
Department of Horticultural Sciences, Cornell University, Geneva, New York 14456.
Smith
F D
FD
Harpending
P R
PR
Rasmussen
J L
JL
Sanford
J C
JC
D001410:Bacillus megaterium / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D005821:Genetic Techniques; D010957:Plasmids; D014170:Transformation, Genetic
D004269:DNA, Bacterial