The nucleotide sequence of a 4.5-kilobase copper resistance determinant from Pseudomonas syringae pv. tomato revealed four open reading frames (ORFs) in the same orientation. Deletion and site-specific mutational analyses indicated that the first two ORFs were essential for copper resistance; the last two ORFs were required for full resistance, but low-level resistance could be conferred in their absence. Five highly conserved, direct 24-base repeats were found near the beginning of the second ORF, and a similar, but less conserved, repeated region was found in the middle of the first ORF.
Cupric sulfate induced mRNA specific to the copper resistance gene cluster previously cloned from Pseudomonas syringae pv. tomato PT23. mRNA from each of the four genes of this cluster responded in a similar manner to induction over time and with different concentrations of cupric sulfate. Promoter fusion constructs indicated the presence of a single copper-inducible promoter upstream from the first open reading frame.
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