Transposon mutagenesis in Acinetobacter calcoaceticus RAG-1.
Molecular genetic studies of Acinetobacter spp. have been greatly limited by the lack of a method for transposon mutagenesis. In this study, a genetically engineered derivative of Tn10, mini-Tn10PttKm, was conjugally transferred in plate matings from Escherichia coli SM10[(lambda pir)(pLOFPttKm)] to Acinetobacter calcoaceticus RAG-1. Transfer frequencies were dependent on mating ratios and varied from 7.9 x 10(-8) to 3.4 x 10(-7) per recipient cell. The 27 lipase-deficient transconjugants which were isolated exhibited several different phenotypes, including gelatinase mutants, esterase mutants, and putative auxotrophs. Southern blot analysis confirmed the insertion of the mini-Tn10PttKm transposon in single, unique sites in five transconjugants. Four of five lipase mutants contained single insertions of mini-Tn10PttKm in the same chromosomal restriction fragments. To our knowledge, this is the first report of the use of a transposon for direct, generalized mutagenesis in Acinetobacter spp.[1]References
- Transposon mutagenesis in Acinetobacter calcoaceticus RAG-1. Leahy, J.G., Jones-Meehan, J.M., Pullias, E.L., Colwell, R.R. J. Bacteriol. (1993) [Pubmed]
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