RANDOM INSERTION OF TRANSPOSON TN4001 IN THE GENOME OF SPIROPLASMA-CITRI STRAIN GII3

Electroporation of Spiroplasma citri strain GII3 with plasmid pMUT con taining the Staphylococcus aureus transposon Tn4001 resulted in random insertion of Tn4001 into the spiroplasmal genome. Transformation freq uencies reached 10(-8) per colony-forming unit (CFU) when 100 mu g 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 mos t cases Tn4001 was randomly inserted in the genome of S. citri strain GII3, without insertion of the carrier plasmid. For most transformed s piroplasmas, Tn4001 was stably maintained in the absence of antibiotic selection for at least 80 bacterial generations, making Tn4001 a pote ntial tool for S. citri mutagenesis. (C) 1997 Academic Press.