Abstract
We have obtained a set ofEscherichia coli K-12 derivatives with transposon-generated Tn10 insertion mutations at thearo genes of their aromatic biosynthetic pathway. Bacteriophage λNK561 (Tn10) has been used for transposon mutagenesis ofE. coli, strain BW545. Tetracycline (Tc)-resistant derivatives were screened by their Aro− phenotype by growth on a minimal medium with adequate requirements. Sixaro mutant types were mapped; two strains werearoA, twoaroD, onearoB oraroE, and onearoC. A selective medium and ad-cycloserine enrichment in the presence of tetracycline were used to select for Aro−, Tc-sensitive derivatives. The reversion index to aromatic-independent colonies of some derivatives was less than 2 × 10−11 per bacterium per generation. P1 transduction experiments transferred an aroA::Tn10 insertion fromE. coli BW545 to an enterotoxigenicE. coli strain from porcine origin. Derivatives of this strain beingaro, Tc-sensitive and not reverting toaro + at a detectable frequency, and many others transduced at will, may prove their usefulness as live vaccines.
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Cobos, A., Fernández, M.F., Hernández, P.E. et al. Transposon-generated Tn10 insertion mutations at thearo genes ofEscherichia coli K-12. Current Microbiology 20, 13–18 (1990). https://doi.org/10.1007/BF02094017
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DOI: https://doi.org/10.1007/BF02094017