FIG. 1.

Physical arrangement of the dca-pca-qui-pob-hca genetic cluster of Acinetobacter sp. strain ADP1 and the strategy used to capture the hca genes. (A) Organization of the genetic regions surrounding the pca gene cluster in the wild-type strain. (B) Introduction of an NdeI deletion that removed DNA from an intergenic dca region up to the pobS gene to form strain ADP8010, defective in adipate, protocatechuate, and quinate catabolism. The small segment of DNA on the left of some NdeI sites (▤) is part of a dca intergenic region. (C) Transformation of strain ADP8010 by pZR8038, followed by selection on adipate, yielding heterogenotes in which a single crossover restored the ability of cells to utilize adipate. MCS, multiple cloning site. (D) Digestion of a lysate of the heterogenote ADP8010::pZR8038 with XbaI followed by ligation and transformation of E. coli, yielding plasmid pZR8200, which was found to have a 16.5-kb insert containing all of the pob and hca genes.