PMID:1991710

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
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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
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