1887

Abstract

Attempts to construct strains with a defect in the common aromatic amino acid biosynthesis pathway were made. In other bacteria the genes of this pathway () can be disrupted in the presence of suitable media supplements. The genomic organization of the genes in reveals that there is one operon () and three isolated genes (, and ). The gene was chosen as a target for disruption; this encodes shikimate kinase, which catalyses the fifth step in chorismate biosynthesis. Attempts to replace the wild-type gene with a disrupted allele (Δ::) by a two-step homologous recombination procedure were unsuccessful in a wild-type strain. When a second functional copy of was integrated into the chromosome, it was possible to isolate a strain carrying the disrupted gene. Excision of the L5-integrated copy of by the L5 excisionase could be not be achieved in the strain carrying the disrupted copy, but was possible in a strain carrying a wild-type copy. These results demonstrate that the chorismate pathway is essential for the viability of

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2002-10-01
2020-11-25
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