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Construction of arsB and tetH mutants of the sulfur-oxidizing bacterium Acidithiobacillus caldus by marker exchange

dc.contributor.authorVan Zyl L.J.
dc.contributor.authorVan Munster J.M.
dc.contributor.authorRawlings D.E.
dc.date.accessioned2011-05-15T15:56:31Z
dc.date.available2011-05-15T15:56:31Z
dc.date.issued2008
dc.identifier.citationApplied and Environmental Microbiology
dc.identifier.citation74
dc.identifier.citation18
dc.identifier.issn992240
dc.identifier.other10.1128/AEM.01235-08
dc.identifier.urihttp://hdl.handle.net/10019.1/9898
dc.description.abstractAcidithiobacillus caldus is a moderately thermophilic, acidophilic bacterium that has been reported to be the dominant sulfur oxidizer in stirred-tank processes used to treat gold-bearing arsenopyrite ores. It is also widely distributed in heap reactors used for the extraction of metals from ores. Not only are these bacteria commercially important, they have an interesting physiology, the study of which has been restricted by the nonavailability of defined mutants. A recently reported conjugation system based on the broad-host-range IncW Plasmids pSa and R388 was used to transfer mobilizable narrow-host-range suicide plasmid vectors containing inactivated and partially deleted chromosomal genes from Escherichia coli to A. caldus. Through the dual use of a selectable kanamycin resistance gene and a hybridization probe made from a deleted portion of the target chromosomal gene, single- and double-recombinant mutants of A. caldus were isolated. The functionality of the gene inactivation system was shown by the construction of A. caldus arsB and tetH mutants, and the effects of these mutations on cell growth in the presence of arsenic and by means of tetrathionate oxidation were demonstrated. Copyright © 2008, American Society for Microbiology. All Rights Reserved.
dc.subjectArsenic
dc.subjectArsenic compounds
dc.subjectCell growth
dc.subjectEscherichia coli
dc.subjectExtraction
dc.subjectGold
dc.subjectGold deposits
dc.subjectGrowth kinetics
dc.subjectHealth
dc.subjectMetal recovery
dc.subjectMetals
dc.subjectMinerals
dc.subjectNonmetals
dc.subjectOres
dc.subjectSulfur
dc.subjectAcidithiobacillus caldus
dc.subjectConjugation systems
dc.subjectDual use
dc.subjectGene inactivation
dc.subjectHybridization probe
dc.subjectKanamycin resistance
dc.subjectModerately thermophilic
dc.subjectPlasmid vectors
dc.subjectSulfur-oxidizing bacterium
dc.subjectTetrathionate
dc.subjectOre treatment
dc.subjectarsenic
dc.subjectarsenopyrite
dc.subjectchromosome
dc.subjectextraction
dc.subjectgenetic marker
dc.subjectgrowth response
dc.subjecthost range
dc.subjecthybridization
dc.subjectmutation
dc.subjectoxidation
dc.subjectphysiology
dc.subjectplasmid
dc.subjectsulfur
dc.subjectthermophilic bacterium
dc.subjectAcidithiobacillus
dc.subjectarticle
dc.subjectbacterium conjugation
dc.subjectbacterium mutant
dc.subjectcell growth
dc.subjectDNA hybridization
dc.subjectgene deletion
dc.subjectgene inactivation
dc.subjectnonhuman
dc.subjectoxidation
dc.subjectsulfur oxidizing bacterium
dc.subjectbacterial chromosome
dc.subjectbacterial gene
dc.subjectdrug effect
dc.subjectEscherichia coli
dc.subjectgene targeting
dc.subjectgene vector
dc.subjectgenetic marker
dc.subjectgenetics
dc.subjectgrowth, development and aging
dc.subjectmetabolism
dc.subjectmutagenesis
dc.subjectplasmid
dc.subjectAcidithiobacillus caldus
dc.subjectBacteria (microorganisms)
dc.subjectEscherichia coli
dc.subjectarsenic trioxide
dc.subjectarsenous acid derivative
dc.subjectbacterial DNA
dc.subjecttetrathionic acid
dc.subjectAcidithiobacillus
dc.subjectArsenites
dc.subjectChromosomes, Bacterial
dc.subjectConjugation, Genetic
dc.subjectDNA, Bacterial
dc.subjectEscherichia coli
dc.subjectGene Targeting
dc.subjectGenes, Bacterial
dc.subjectGenetic Markers
dc.subjectGenetic Vectors
dc.subjectMutagenesis
dc.subjectPlasmids
dc.subjectTetrathionic Acid
dc.titleConstruction of arsB and tetH mutants of the sulfur-oxidizing bacterium Acidithiobacillus caldus by marker exchange
dc.typeArticle
dc.description.versionArticle


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