Development of a plasmid transfer system for the anaerobic sulphate reducer, Desulfovibrio vulgaris

Berg, W.A.M. van den; Stokkermans, J.P.W.G.; Dongen, W.M.A.M. van

doi:10.1016/0168-1656(89)90014-x

Cited by 31 publications

(16 citation statements)

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“…Unfortunately, the genetics of the sulfate-reducing bacteria have lagged far behind their physiological and biochemical characterization (Wall, 1993). Conjugal transfer of broad-host-range plasmids belonging to iiicompatibility group IncQ is presently the only gene transfer method available for D. vulgaris (Argyle e t al., 1992;Powell et al, 1989;van den Berg et al, 1989).…”

Section: R F U a N D G V O O R D O U Wmentioning

confidence: 99%

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Targeted gene-replacement mutagenesis of dcrA, encoding an oxygen sensor of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough

Voordouw

1997

Microbiology

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A gene-replacement mutagenesis method has been developed for the anaerobic, sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough and used to delete dcrA, encoding a potential oxygen or redox sensor with homology to the methyl-accepting chemotaxis proteins. A suicide plasmid, containing a cat-marked dcrA allele and a counter-selectable sacB marker was transferred from Escherichia coli 517-1 to D. vulgaris by conjugation. Following plasmid integration the desired dcrA deletion mutant (D. vulgaris F100) was obtained in media containing sucrose and chloramphenicol. Southern blot screening was required to distinguish D. vulgaris FIOO from strains in which the sacB marker was inactivated by transposition of an endogenous IS element. No anaerotactic deficiency has so far been detected in D. vulgaris F100, which was found t o be more resistant to inactivation by oxygen than the wild-type. Increased transcription of the rho-rub operon, located immediately downstream from dcrA, 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 E. coli.

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Section: R F U a N D G V O O R D O U Wmentioning

confidence: 99%

“…Attempts to use conjugation of suicide plasmids for construction of specific gene mutants of D. vulgaris were unsuccessful (van den Berg et al, 1989;van Dongen et a/., 1994). The intriguing biochemical properties of DcrA provided a strong incentive to revisit the problem of gene interruption or replacement in D. vulgaris.…”

Section: R F U a N D G V O O R D O U Wmentioning

confidence: 99%

Targeted gene-replacement mutagenesis of dcrA, encoding an oxygen sensor of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough

Voordouw

1997

Microbiology

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“…Finally, the six-electron reduction of sulfite to sulfide (E 0 = ϭ Ϫ116 mV) is carried out by dissimilatory sulfite reductase (equation 3) and is apparently coupled to electron transport-linked phosphorylation. Although the immediate electron donor for this reaction is not proven, electrons from H 2 can support this reduction.Members of the genus Desulfovibrio have been widely used as experimental models for biochemical and molecular studies of sulfate reduction because these strains have been easy to cultivate and were the first to be genetically manipulated (7,8). Desulfovibrio strains incompletely oxidize the preferred substrates lactate and pyruvate (9-11), as shown in equation 4 for lactate.…”

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confidence: 99%

“…Members of the genus Desulfovibrio have been widely used as experimental models for biochemical and molecular studies of sulfate reduction because these strains have been easy to cultivate and were the first to be genetically manipulated (7,8). Desulfovibrio strains incompletely oxidize the preferred substrates lactate and pyruvate (9-11), as shown in equation 4 for lactate.…”

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New Model for Electron Flow for Sulfate Reduction in Desulfovibrio alaskensis G20

Keller

Rapp-Giles

Semkiw

et al. 2014

Appl Environ Microbiol

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To understand the energy conversion activities of the anaerobic sulfate-reducing bacteria, it is necessary to identify the components involved in electron flow. The importance of the abundant type I tetraheme cytochrome c 3 (TpIc 3 ) as an electron carrier during sulfate respiration was questioned by the previous isolation of a null mutation in the gene encoding TpIc 3 , cycA, in Desulfovibrio alaskensis G20. Whereas respiratory growth of the CycA mutant with lactate and sulfate was little affected, growth with pyruvate and sulfate was significantly impaired. We have explored the phenotype of the CycA mutant through physiological tests and transcriptomic and proteomic analyses. Data reported here show that electrons from pyruvate oxidation do not reach adenylyl sulfate reductase, the enzyme catalyzing the first redox reaction during sulfate reduction, in the absence of either CycA or the type I cytochrome c 3 :menaquinone oxidoreductase transmembrane complex, QrcABCD. In contrast to the wild type, the CycA and QrcA mutants did not grow with H 2 or formate and sulfate as the electron acceptor. Transcriptomic and proteomic analyses of the CycA mutant showed that transcripts and enzymes for the pathway from pyruvate to succinate were strongly decreased in the CycA mutant regardless of the growth mode. Neither the CycA nor the QrcA mutant grew on fumarate alone, consistent with the omics results and a redox regulation of gene expression. We conclude that TpIc 3 and the Qrc complex are D. alaskensis components essential for the transfer of electrons released in the periplasm to reach the cytoplasmic adenylyl sulfate reductase and present a model that may explain the CycA phenotype through confurcation of electrons. E nvironmental impacts of the sulfate-reducing bacteria (SRB), such as the formation of toxic sulfide and corrosion of metals, stem from SRB sulfate respiration and from vigorous metal metabolism, including both reduction and oxidation. The involvement of these bacteria in microbially influenced metal corrosion (1, 2) and their possible application for the remediation of toxic metal-contaminated environments (3, 4) have driven a systems biology investigation of their metabolism. To predict or control metabolic capabilities for beneficial environmental purposes, the bioenergetic pathways of the SRB need to be elucidated in more detail.Typically, SRB of the genus Desulfovibrio use H 2 , organic acid substrates, formate, or short-chain alcohols as electron donors for sulfate reduction, a process that occurs in the cytoplasm and is mediated by soluble enzymes. With hydrogen as the source of electrons, at an environmentally relevant partial pressure of 10 Pa (reduction potential [E=] ϭ Ϫ300 mV) (5), the first two-electron reduction of sulfate to sulfite (E 0 = ϭ Ϫ516 mV) cannot be accomplished without an additional energy input. Sulfate is activated at the expense of two ATP equivalents through the activity of sulfate adenylyltransferase (equation 1), producing adenosine phosphosulfate (APS), which increases th...

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“…The amino acid sequence (107 residues) of mature cytochrome C3 from Desulfovibrio vulgaris subsp. vulgaris Hildenborough (referred to hereafter as D. vulgaris Hildenborough) (26,30; R. P. Ambler, Biochem. J.…”

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Functional expression of Desulfovibrio vulgaris Hildenborough cytochrome c3 in Desulfovibrio desulfuricans G200 after conjugational gene transfer from Escherichia coli

et al. 1990

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Plasmid pJRDC800-1, containing the cyc gene encoding cytochrome c3 from Desulfovibrio vulgaris subsp. vulgaris Hildenborough, was transferred by conjugation from Escherichia coli DH5 alpha to Desulfovibrio desulfuricans G200. The G200 strain produced an acidic cytochrome c3 (pI = 5.8), which could be readily separated from the Hildenborough cytochrome c3 (pI = 10.5). The latter was indistinguishable from cytochrome c3 produced by D. vulgaris subsp. vulgaris Hildenborough with respect to a number of chemical and physical criteria.

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Development of a plasmid transfer system for the anaerobic sulphate reducer, Desulfovibrio vulgaris

Cited by 31 publications

References 20 publications

Targeted gene-replacement mutagenesis of dcrA, encoding an oxygen sensor of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough

Targeted gene-replacement mutagenesis of dcrA, encoding an oxygen sensor of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough

New Model for Electron Flow for Sulfate Reduction in Desulfovibrio alaskensis G20

Functional expression of Desulfovibrio vulgaris Hildenborough cytochrome c3 in Desulfovibrio desulfuricans G200 after conjugational gene transfer from Escherichia coli

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