Foissac X, Saillard C, Bové J M
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.
Plasmid. 1997;37(1):80-6. doi: 10.1006/plas.1996.1271.
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.
用含有金黄色葡萄球菌转座子Tn4001的质粒pMUT对柑橘螺原体菌株GII3进行电穿孔，导致Tn4001随机插入螺原体基因组。当使用100微克质粒DNA和3×10⁹个柑橘螺原体CFU时，转化频率达到每菌落形成单位（CFU）10⁻⁸。其他三株柑橘螺原体在相同条件下未能被转化。在大多数情况下，Tn4001随机插入柑橘螺原体菌株GII3的基因组中，而载体质粒未插入。对于大多数转化的螺原体，在没有抗生素选择的情况下，Tn4001至少在80个细菌世代中稳定维持，这使得Tn4001成为柑橘螺原体诱变的潜在工具。
