IS1491fromPseudomonas alcaligenesNCIB 9867: Characterization and Distribution amongPseudomonasSpecies

A new insertion sequence, IS1491,has been cloned and sequenced. The 2489-bp IS1491was isolated from aPseudomonas alcaligenesNCIB 9867 (strain P25X) 4.8-kbPstI chromosomal fragment. IS1491is flanked by an imperfect inverted repeat of 23 bp and carries two overlapping open reading frames, ORF1 and ORF2. Both ORF1 and ORF2 displayed homology to the IstA-like and IstB-like transposases encoded by the IS21family of insertion sequences, which include two IS elements previously isolated fromP. alcaligenesP25X, IS1474,and IS1475(Yeo, C. C., and Poh, C. L. (1997).FEMS Microbiol. Lett.149,257–263). Transposition assays showed that IS1491transposed at a frequency of approximately 1.4 × 10⁻⁶. Transposition of IS1491into the target pRK415 replicon was observed but when ORF2 was disrupted, a fusion between the donor and target replicons was detected. IS1491-like sequences were detected in total DNA ofPseudomonas putidaNCIB 9869 (strain P35X),Pseudomonas aeruginosa, Pseudomonas stutzeri, Pseudomonas syringae, Pseudomonas mendocina, Comomonas acidovorans,andComomonas testosteroniby hybridization with IS1491DNA.