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 μg plasmid DNA and 107-108M. 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. © 1993 Academic Press. All rights reserved.