Construction of an unmarked mutant of ataA from Acinetobacter sp. Tol 5. (A) Genetic organization around ataA in Acinetobacter sp. Tol 5 and its derivative mutants obtained by plasmid integration and FLP/FRT recombination. The arrows indicate the primers used in PCR analysis for the confirmation of the constructs. (B) PCR confirmation of plasmid integration and the deletion of ataA in the Tol 5 derivatives. Chromosomal DNA was extracted as a template for PCR from Tol 5 and its derivatives (G4, G4K1, and 4140). PCR analyses were performed by using three different primer sets: P1 (AtaAupstF2) + P2 (FRT-SP6R), P3 (FRT-leftF) + P4 (AtaAdwstR2), and P1 + P4. The nucleotide sequences of these primers are shown in Table 2.