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Volume 143, Issue 6

Targeted gene-replacement mutagenesis of encoding an oxygen sensor of the sulfate-reducing bacterium Hildenborough Free

Abstract

A gene-replacement mutagenesis method has been developed for the anaerobic, sulfate-reducing bacterium Hildenborough and used to delete encoding a potential oxygen or redox sensor with homology to the methyl-accepting chemotaxis proteins. A suicide plasmid, containing a -marked allele and a counter-selectable marker was transferred from S17-1 to by conjugation. Following plasmid integration the desired deletion mutant ( F100) was obtained in media containing sucrose and chloramphenicol. Southern blot screening was required to distinguish F100 from strains in which the marker was inactivated by transposition of an endogenous IS element. No anaerotactic deficiency has so far been detected in F100, which was found to be more resistant to inactivation by oxygen than the wild-type. Increased transcription of the operon, located immediately downstream from was demonstrated by Northern blotting and may be the cause of this unusual phenotype, in view of the recent discovery that Rbo can complement the deleterious effects of superoxide dismutase deficiency in

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Keyword(s): aerotaxis , Desulfovibrio , gene replacement , oxygen sensor and superoxide dismutase
© Society Society for General Microbiology 1997
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1997-06-01
2024-04-26
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Targeted gene-replacement mutagenesis of dcrA, encoding an oxygen sensor of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough
Microbiology 143, 1815 (1997); https://doi.org/10.1099/00221287-143-6-1815
/content/journal/micro/10.1099/00221287-143-6-1815
/content/journal/micro/10.1099/00221287-143-6-1815
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