PMID:4933690

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Citation

Wilson, TH, Kusch, M and Kashket, ER (1970) A mutant in Escherichia coli energy-uncoupled for lactose transporta defect in the lactose-operon.Biochem. Biophys. Res. Commun. 40:1409-14

Abstract

No abstract in PubMed

Links

PubMed

Keywords

Biological Transport, Active; Carbon Isotopes; Cell Membrane; Coliphages; Escherichia coli; Galactose; Glycosides; Lactose; Molecular Biology; Mutation; Nitrobenzenes; Operon; Transduction, Genetic

Main Points of the Paper

  • Isolation of mutant (X71-54) with a severe defect in TMG accumulation
    • possesses elevated membrane carrier activity, therefore the rate of efflux is increased

Materials and Methods Used

  • Parent Strain: X71 (i-z+y+a-, proC-, try-, B1-, F-)
  • Additional Strain: X71-54
  • Additional Strain: Ca77 (lactose-deleted, B1-, Hfr)
  • β-galactosidase assay
  • ONPG uptake assay
  • Thiomethylgalactoside (TMG) transport assay
  • P1 Transduction

Phenotype Annotations

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Species Taxon ID Strain Gene (if known) OMP Phenotype Details Evidence Notes

Escherichia coli

NCBI:83333

X71-54

lacYUN

  • Decreased ability to maintain concentration gradient?
  • Abolished energy-coupling transporter activity?
  • Absent intracellular accumulation of thiomethylgalactoside (direct observation)

Other

Intracellular [14C]TMG accumulation

Other

Figure 1- Filter assay

Escherichia coli

NCBI:83333

X71-54

lacYUN

  • Decreased intracellular accumulation of substance
  • Increased rate of transporter efflux?
  • Decreased energy-coupled transport?
  • Increased rate of efflux of thiomethylgalactoside (TMG) (direct observation)

Other

exit rate of [14C]TMG was faster than that found for the parent

Other

Figure 2- Filter assay

Escherichia coli

NCBI:83333

X71-54

lacYUN

  • Increased rate of transporter substrate uptake?
  • Increased transport of small molecules
  • Increased rate of transporter activity

Metabolic Activity

130% of normal uptake activity (gauged by ONPG entry)

Biochemical Assay

Table 1- ONPG assay

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Notes

Additional papers regarding this mutant can be found here