Disruption of the gene of mc155, by standard gene replacement methods, revealed that there are two copies of this gene within a large duplication of the mc155 genome. AdhC and AdhC mutants were obtained when one or two copies, respectively, were disrupted by homologous recombination. Southern blot analysis of I restriction digests of the DNA from these mutants and from wild-type mc155, resolved by PFGE, showed that the duplication size may be at least ∼250 kb. The single and double knockout mutants were characterized and compared with the wild-type. A growth disadvantage and a different morphology were associated with the loss of expression of one or both of the copies, but both mutants were still acid-fast. Findings in this study indicate that the process of chromosomal duplication in is ongoing and remains a potent source of genome dynamics. Hence, the mc155 genome might be larger than previously thought.


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