FIG. 1.

Genetic organization of the cloned region including ald1 and that of disrupted ald1. (A) Restriction map of the 5.7-kb HindIII cloned fragment. (B) The ORFs within the sequenced region of the 3.5-kb PstI fragment. ORF1 showed homology with a hypothetical 47.3-kDa protein in the thcA 5′ region (ORF3) from Rhodococcus sp. strain NI86/21 (GenBank accession number U17129) (15), and ORF2 showed homology with ethanolamine permease from Rhodococcus sp. strain NI86/21 (GenBank accession number L24492) (4). (C) Construction of the gene disruption vector pDALD1, derived from pSH6. The hatched box represents the kanamycin resistance gene. (D) Genomic Southern analysis of EcoRV-KpnI double-digested total DNAs (5.0 μg each) from the wild-type strain (lane 1) and the ald1Δ strain (lane 2), with the ³²P-labeled ald1 fragment as a probe.