Random insertion of the gentamicin resistance transposon Tn4001 in Mycoplasma pulmonis.
The staphylococcal transposon Tn4001 was introduced into Mycoplasma pulmonis using an Escherichia coli-derived vector by polyethylene glycol-mediated transformation. Using a reaction mixture containing 10 micrograms plasmid DNA, 10 micrograms yeast tRNA, and 34-35% polyethylene glycol per 1 x 10(8) cells, Tn4001 could be introduced into M. pulmonis at a frequency of 5 x 10(-5) per colony forming unit. DNA-DNA hybridization studies illustrated that Tn4001 could occupy a diversity of insertion sites in the M. pulmonis chromosome. These data indicated that Tn4001 is a potentially useful tool for the introduction of mutations and for genetic studies in M. pulmonis.
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Veterinary Medical Research Institute, College of Veterinary Medicine, Iowa State University, Ames 50011.
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D015139:Blotting, Southern; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D004352:Drug Resistance, Microbial / Q000235:genetics; D005839:Gentamicins / Q000494:pharmacology*; D009174:Mycoplasma / Q000187:drug effects / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D011815:R Factors; D015183:Restriction Mapping
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D005839:Gentamicins