1887

Abstract

Phages capable of transducing succinate dehydrogenase mutants () of were isolated from pools of artificially constructed recombinant lambda phages using a selective casein digest medium. These phages produced characteristically dense turbid plaques, and as prophages they increased the aerobic growth efficiencies of mutants on complex media but were unable to promote growth with succinate as sole carbon and energy source (an essential feature of strains). The phages were identified as fumarate reductase transducing phages (λ) by the presence of a characteristic 4·9 kilobase pairs R. dIII fragment of bacterial DNA, the expression of a polypeptide with a relative molecular mass of 72000 (the gene product) and by comparing their transducing activities with authentic λ phages. In parallel studies a strain containing a ColE1- hybrid plasmid (pGS1 = pLC16.43) was characterized. Transfer of pGS1 to mutants was accompanied by increased aerobic growth efficiencies on complex media and the ability to utilize succinate as sole carbon and energy source. It was concluded that fumarate reductase can replace succinate dehydrogenase but the extent of the reversal of the lesion depends on gene dosage and the titration of the repressor which normally prevents aerobic synthesis of the reductase.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-122-2-171
1981-02-01
2021-08-03
Loading full text...

Full text loading...

/deliver/fulltext/micro/122/2/mic-122-2-171.html?itemId=/content/journal/micro/10.1099/00221287-122-2-171&mimeType=html&fmt=ahah

References

  1. Borck K., Beggs J.D., Brammar W.J., Hopkins A.S., Murray N.E. 1976; The construction in vitro of transducing derivatives of phage lambda. Molecular and General Genetics 146:199–207
    [Google Scholar]
  2. Clarke L., Carbon J. 1976; A colony bank containing synthetic ColEl hybrid plasmids representative of the entire E. coli genome. Cell 9:91–99
    [Google Scholar]
  3. Cole S.T., Guest J.R. 1978; Studies with artificially-constructed fumarate reductase transducing phages (λrdA) . Society for General Microbiology Quarterly 6:42–43
    [Google Scholar]
  4. Cole S.T., Guest J.R. 1979a; Amplification and aerobic synthesis of fumarate reductase in ampicillin-resistant mutants of Escherichia coli K. 12. FEMS Microbiology Letters 5:65–67
    [Google Scholar]
  5. Cole S.T., Guest J.R. 1979b; Production of a soluble form of fumarate reductase by multiple gene duplication in Escherichia coli K12. European Journal of Biochemistry 102:65–71
    [Google Scholar]
  6. Cole S.T., Guest J.R. 1980a; Genetic and physical characterization of lambda transducing phages (λfrdA) containing the fumarate reductase gene of Escherichia coli K12. Molecular and General Genetics 178:409–418
    [Google Scholar]
  7. Cole S.T., Guest J.R. 1980b; Amplification of fumarate reductase synthesis with λfrdA transducing phages and orientation of frdA gene expression. Molecular and General Genetics 179:377–385
    [Google Scholar]
  8. Creaghan I.T., Guest J.R. 1978; Succinate dehydrogenase-dependent nutritional requirement for succinate in mutants of Escherichia coli K12. Journal of General Microbiology 107:1–13
    [Google Scholar]
  9. Edlund T., Grundström T., Normark S. 1979; Isolation and characterization of DNA repetitions carrying the chromosomal β-lactamase gene of Escherichia coli K-12. Molecular and General Genetics 173:115–125
    [Google Scholar]
  10. Guest J.R., Stephens P.E. 1980; Molecular cloning of the pyruvate dehydrogenase complex genes of Escherichia coli . Journal of General Microbiology 121:277–292
    [Google Scholar]
  11. Hirsch C.A., Rasminsky M., Davis B.D., Lin E.C.C. 1963; A fumarate reductase in Escherichia coli distinct from succinate dehydrogenase. Journal of Biological Chemistry 238:3770–3774
    [Google Scholar]
  12. Ruch F.E., Kuritzkes D.R., Lin E.C.C. 1979; Use of lac operon fusions to isolate Escherichia coli mutants with altered expression of the fumarate reductase system in response to substrate and respiratory controls. Biochemical and Biophysical Research Communications 91:1365–1370
    [Google Scholar]
  13. Spencer M.E., Guest J.R. 1973; Isolation and properties of fumarate reductase mutants of Escherichia coli . Journal of Bacteriology 114:563–570
    [Google Scholar]
  14. Spencer M.E., Guest J.R. 1974; Proteins of the inner membrane of Escherichia coli: changes in composition associated with anaerobic growth and fumarate reductase amber mutation. Journal of Bacteriology 117:954–959
    [Google Scholar]
  15. Wilson G.G., Murray N.M. 1979; Molecular cloning of the DNA ligase gene from bacteriophage T4. I. Characterisation of the recombinants. Journal of Molecular Biology 132:471–491
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-122-2-171
Loading
/content/journal/micro/10.1099/00221287-122-2-171
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error