Reversal by DNA amplifications of an unusual mutation blocking alkane and alcohol utilization in Pseudomonas putida

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Abstract
We analyzed the reversion of strains carrying alk208, a mutation in the alkBAC (alkane utilization) region of the Pseudomonas CAM-OCT plasmid. Reversion of alk208 was stimulated 25 to 75-fold by small doses of UV-irradiation. All alkane hydroxylase-positive (AlkB+) revertants proved to be aliphatic alcohol dehydrogenasepositive (AlkC+) as well, whereas AlkC+ revertants could be either AlkB+ or AlkB-. Most of the AlkB- AlkC+ partial revertants produced AlkC- segregants at measurable frequencies. UV-irradiation substantially increased the rate of AlkC- segregation. Most segregants reverted to AlkB+ or AlkC+ at frequencies similar to the original alk208 strain. Dot blot hybridization analyses using cloned probes from various regions of CAM-OCT revealed that the partial revertants contained specific amplications of alk DNA. The endpoints of these amplifications mapped in at least two regions. AlkC- segregants had lost the DNA amplifications.