PMID:1991710

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Citation

Santangelo, JD, Jones, DT and Woods, DR (1991) Metronidazole activation and isolation of Clostridium acetobutylicum electron transport genes.J. Bacteriol. 173:1088-95

Abstract

An Escherichia coli F19 recA, nitrate reductase-deficient mutant was constructed by transposon mutagenesis and shown to be resistant to metronidazole. This mutant was a most suitable host for the isolation of Clostridium acetobutylicum genes on recombinant plasmids, which activated metronidazole and rendered the E. coli F19 strain sensitive to metronidazole. Twenty-five E. coli F19 clones containing different recombinant plasmids were isolated and classified into five groups on the basis of their sensitivity to metronidazole. The clones were tested for nitrate reductase, pyruvate-ferredoxin oxidoreductase, and hydrogenase activities. DNA hybridization and restriction endonuclease mapping revealed that four of the C. acetobutylicum insert DNA fragments on recombinant plasmids were linked in an 11.1-kb chromosomal fragment. DNA sequencing and amino acid homology studies indicated that this DNA fragment contained a flavodoxin gene which encoded a protein of 160 amino acids that activated metronidazole and made the E. coli F19 mutant very sensitive to metronidazole. The flavodoxin and hydrogenase genes which are involved in electron transfer systems were linked on the 11.1-kb DNA fragment from C. acetobutylicum.

Links

PubMed

Keywords

Amino Acid Sequence; Base Sequence; Blotting, Southern; Clostridium; Electron Transport; Escherichia coli; Flavodoxin; Gene Expression Regulation, Bacterial; Gene Library; Genes, Bacterial; Genetic Linkage; Hydrogenase; Ketone Oxidoreductases; Metronidazole; Molecular Sequence Data; Mutagenesis, Insertional; Nitrate Reductase; Nitrate Reductases; Pyruvate Synthase; Restriction Mapping; Sequence Homology, Nucleic Acid

Main Points of the Paper

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Materials and Methods Used

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Phenotype Annotations

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Phenotype of Taxon Information Genotype Information (if known) Condition Information OMP ID OMP Term Name ECO ID ECO Term Name Notes Status

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: F19
  • NCBI Taxon ID: 8333
  • Genotype of Reference Strain: recA
  • Genotype of Experimental Strain : recA
  • Reference Condition:
OMP:0000276

increased resistance to antibiotics

ECO:0000182

in vitro culture assay data

Resistance to Metronidazole, figure 1

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: P460
  • NCBI Taxon ID: 8333
  • Genotype of Reference Strain: ompA
  • Genotype of Experimental Strain : ompA
  • Reference Condition:
OMP:0000276

increased resistance to antibiotics

ECO:0000182

in vitro culture assay data

Resistant to bacteriocin Colicin K, table 3

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: P1636
  • NCBI Taxon ID: 8333
  • Genotype of Reference Strain: ompA
  • Genotype of Experimental Strain : ompA
  • Reference Condition:
OMP:0000276

increased resistance to antibiotics

ECO:0000182

in vitro culture assay data

Resistant to bacteriocin Colicin L, table 3

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: P460
  • NCBI Taxon ID: 8333
  • Genotype of Reference Strain: ompA
  • Genotype of Experimental Strain : ompA
  • Reference Condition:

OMP:0000038

bacteriophage-host interaction phenotype

ECO:0000182

in vitro culture assay data

Resistant to bacteriophage K3


Notes

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