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Genetic and physical characterization of lambda transducing phages (λfrdA) containing the fumarate reductase gene of Escherichia coli K12

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Summary

Two types of fumarate reductase transducing phages, λfrdA, carrying the wild-type frdA gene but differing in the orientation of a R.HindIII fragment of bacterial DNA were isolated from populations of recombinant transducing phages by their ability to complement the lesions of frdA mutants of E. coli. In lysogens, the cloned frdA gene was controlled by its own promoter and was fully responsive to normal regulatory stimuli. The λfrdA phages would not complement the defects of succinate dehydrogenase (sdh) mutants. Genetic studies showed that the R.HindIII fragment contains ampA, the cis-acting regulatory locus for the chromosomal β-lactamase gene ampC. No evidence for the presence of other markers was detected but the bacterial segment could be extended to produce plaque-forming phage derivatives containing the amp operon and a gene concerned with bacteriophage morphogenesis, groE(mop). A physical map of the 4.9 kb R.HindIII fragment was constructed by restriction analysis and flanking fragments were identified by DNA: DNA hybridization analysis. The frdA region contained a single asymmetric R.EcoRI target 3.33 kb from one end and the orientation of the physical map with respect to the E. coli linkage map was established.

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Authors and Affiliations

  1. Department of Microbiology, University of Sheffield, Western Bank, S10 2TN, Sheffield, England

    Stewart T. Cole & John R. Guest

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  1. Stewart T. Cole
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  2. John R. Guest
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Communicated by D. Sherratt

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Cole, S.T., Guest, J.R. Genetic and physical characterization of lambda transducing phages (λfrdA) containing the fumarate reductase gene of Escherichia coli K12. Molec. Gen. Genet. 178, 409–418 (1980). https://doi.org/10.1007/BF00270492

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