Random insertion of transposon Tn4001 in the genome of Spiroplasma citri strain GII3.
Electroporation of Spiroplasma citri strain GII3 with plasmid pMUT containing the Staphylococcus aureus transposon Tn4001 resulted in random insertion of Tn4001 into the spiroplasmal genome. Transformation frequencies reached 10(-8) per colony-forming unit (CFU) when 100 microg of plasmid DNA and 3 x 10(9) S. citri CFU were used. Three other strains of S. citri failed to be transformed under the same conditions. In most cases Tn4001 was randomly inserted in the genome of S. citri strain GII3, without insertion of the carrier plasmid. For most transformed spiroplasmas, Tn4001 was stably maintained in the absence of antibiotic selection for at least 80 bacterial generations, making Tn4001 a potential tool for S. citri mutagenesis.
Foissac
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Laboratoire de Biologie Cellulaire et Moléculaire, Institut National de la Recherche Agronomique and Université de Bordeaux II, Domaine de la Grande Ferrade, 33883 Villenave d'Ornon cedex, France.
Saillard
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C
Bové
J M
JM
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D004251:DNA Transposable Elements; D004352:Drug Resistance, Microbial / Q000235:genetics; D018274:Electroporation; D004926:Escherichia coli / Q000235:genetics; D016680:Genome, Bacterial; D005839:Gentamicins / Q000494:pharmacology; D009154:Mutation; D013149:Spiroplasma / Q000187:drug effects / Q000235:genetics*; D013211:Staphylococcus aureus / Q000235:genetics; D014170:Transformation, Genetic
D000900:Anti-Bacterial Agents; D004251:DNA Transposable Elements; D005839:Gentamicins