Transformation of Mycoplasma pneumoniae with Tn4001 by electroporation.
Mycoplasma pneumoniae was transformed with the Staphylococcus aureus transposon Tn4001 by electroporation. A transformation frequency of 10(-3) to 10(-5)/colony-forming unit was observed using 30.0 microgram plasmid DNA and 10(7)-10(8) M. pneumoniae colony-forming units. DNA hybridization analyses using standard and pulsed field agarose gel electrophoresis confirmed chromosomal insertion of the transposon, apparently by a transpositional mechanism into random sites. These studies demonstrate the functionality of Tn4001 in M. pneumoniae and suggest its potential as a genetic tool in this mycoplasma.
Hedreyda
C T
CT
Department of Microbiology, University of Georgia, Athens 30602.
Lee
K K
KK
Krause
D C
DC
D015139:Blotting, Southern; D002876:Chromosomes, Bacterial; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004269:DNA, Bacterial / Q000096:biosynthesis / Q000302:isolation & purification; D018274:Electroporation; D016254:Mutagenesis, Insertional / Q000379:methods; D009177:Mycoplasma pneumoniae / Q000235:genetics*; D015183:Restriction Mapping; D013211:Staphylococcus aureus / Q000235:genetics*; D014169:Transformation, Bacterial
D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes