Quantitative evaluation of<i>Escherichia coli</i>host strains for tolerance to cytosine methylation in plasmid and phage recombinants

Woodcock, David M.; Crowther, P.J.; Doherty, Judy P.; Jefferson, Stacie; Decruz, Exmond E.; Noyer-Weidner, Mario; Smith, Steven S.; Michael, Michael; Graham, Michael W.

doi:10.1093/nar/17.9.3469

Cited by 707 publications

(226 citation statements)

References 19 publications

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“…Escherichia coli strain DH5a [supE44 w80lacZDM15 D(lacZYA-argF)U169 deoR recA1 endA1 hsdR17 phoA supE44 thi-11 gyrA96 relA1 l 2 ] (Woodcock et al, 1989) was used for routine DNA manipulations. G. sulfurreducens strain DL1, a wild-type derivative of ATCC 51573 (Caccavo et al, 1994;Coppi et al, 2001), was obtained from our laboratory culture collection.…”

Section: Methodsmentioning

confidence: 99%

Involvement of Geobacter sulfurreducens SfrAB in acetate metabolism rather than intracellular, respiration-linked Fe(III) citrate reduction

et al. 2007

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A soluble ferric reductase, SfrAB, which catalysed the NADPH-dependent reduction of chelated Fe(III), was previously purified from the dissimilatory Fe(III)-reducing micro-organism Geobacter sulfurreducens, suggesting that reduction of chelated forms of Fe(III) might be cytoplasmic. However, metabolically active spheroplast suspensions could not catalyse acetate-dependent Fe(III) citrate reduction, indicating that periplasmic and/or outer-membrane components were required for Fe(III) citrate reduction. Furthermore, phenotypic analysis of an SfrAB knockout mutant suggested that SfrAB was involved in acetate metabolism rather than respiration-linked Fe(III) reduction. The mutant could not grow via the reduction of either Fe(III) citrate or fumarate when acetate was the electron donor but could grow with either acceptor if either hydrogen or formate served as the electron donor. Following prolonged incubation in acetate : fumarate medium in the absence of hydrogen and formate, an 'acetate-adapted' SfrAB-null strain was isolated that was capable of growth on acetate : fumarate medium but not acetate : Fe(III) citrate medium. Comparison of gene expression in this strain with that of the wild-type revealed upregulation of a potential NADPH-dependent ferredoxin oxidoreductase as well as genes involved in energy generation and amino acid uptake, suggesting that NADPH homeostasis and the tricarboxylic acid (TCA) cycle were perturbed in the 'acetate-adapted' SfrAB-null strain. Membrane and soluble fractions prepared from the 'acetate-adapted' strain were depleted of NADPH-dependent Fe(III), viologen and quinone reductase activities. These results indicate that cytoplasmic, respiration-linked reduction of Fe(III) by SfrAB in vivo is unlikely and suggest that deleting SfrAB may interfere with growth via acetate oxidation by interfering with NADP regeneration.

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Section: Methodsmentioning

confidence: 99%

Involvement of Geobacter sulfurreducens SfrAB in acetate metabolism rather than intracellular, respiration-linked Fe(III) citrate reduction

et al. 2007

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“…Bioreactor cultivation was as described previously . Escherichia coli strains DH5aF' (Woodcock et al, 1989) and JMl05 (Pharmacia) were grown in Luria broth. Erythromycin (500 pg ml-l) or ampicillin (25-50 pg ml-l) was added to the growth medium when the pJDC9 and pKK223-3 (Pharmacia) vectors, respectively, were used in E. coli.…”

Section: Methodsmentioning

confidence: 99%

An X-prolyl dipeptidyl aminopeptidase (pepX) gene from Lactobacillus helveticus

Vesanto¹,

Savijoki²,

Rantanen³

et al. 1995

Microbiology

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The X-prolyl dipeptidyl aminopeptidase gene (pepx) of an industrially used Lactobacillus helveticus strain has been detected by nucleic acid hybridization, cloned, characterized and sequenced. One ORF of 2379 bp with coding capacity for a 906 kDa protein (PepX) was found. The ORF was preceded by a typical prokaryotic promoter region. An inverted repeat structure with AG of -84.1 kJ mol-1 was found downstream of the coding region. The deduced amino acid sequence of the 906 kDa protein showed 49*3,494 and 777% homology with the PepX proteins from Lactococcus lactis subsp. lactis, Lc. lactis subsp. cremoris and Ladobacillus delbnreckii subsp. Iactis, respectively. Northern blotting revealed a 2.6 kb transcript and one transcription start site was identified via primer extension analysis using an A.L.F. sequencer. In a bioreactor study, the expression of pepX in Lb. helveticus was studied as a function of growth. Transcription of pepX was typical of exponential growth phase expression. The pepX gene has been cloned into pKK223-3 and expressed at a high level in Escherichia coli JM105. PepX was purified to homogeneity by ion-exchange and hydrophobic interaction chromatography. Optimum PepX activity was observed at pH 6 5 and 45 OC. According to gel filtration analysis, PepX is a dimer of 165 kDa. The enzyme was inactivated by heavy metal ions such as Cu2+, Cd2+ and Zn2+. EDTA and l,l0-phenanthroline did not decrease PepX activity significantly. It was completely inhibited by p-hydroxymercuribenzoate and reactivated by adding Dll, and strongly inhibited by PMSF. PepX is thus a metal-independent serine peptidase having functional sulfhydryl groups at or near the active site.

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“…MATα, SUC2 ) (Boles et al, 1996) was used as the host strain. Escherichia coli strain DH5α (Woodcock et al, 1989) was used as the bacterial cloning host.…”

Section: Strains and Plasmidsmentioning

confidence: 99%

The ORF YNL274c (GOR1) codes for glyoxylate reductase in Saccharomyces cerevisiae

Rintala

Pitkänen

Vehkomäki

et al. 2006

Yeast

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The enzyme glyoxylate reductase reversibly reduces glyoxylate to glycolate, or alternatively hydroxypyruvate to D-glycerate, using either NADPH or NADH as a co-factor. The enzyme has multiple metabolic roles in different organisms. In this paper we show that GOR1 (ORF YNL274c) encodes a glyoxylate reductase and not a hydroxyisocaproate dehydrogenase in Saccharomyces cerevisiae, even though it also has minor activity on α-ketoisocaproate. In addition, we show that deletion of the glyoxylate reductase-encoding gene leads to higher biomass concentration after diauxic shift.

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Quantitative evaluation ofEscherichia colihost strains for tolerance to cytosine methylation in plasmid and phage recombinants

Cited by 707 publications

References 19 publications

Involvement of Geobacter sulfurreducens SfrAB in acetate metabolism rather than intracellular, respiration-linked Fe(III) citrate reduction

Involvement of Geobacter sulfurreducens SfrAB in acetate metabolism rather than intracellular, respiration-linked Fe(III) citrate reduction

An X-prolyl dipeptidyl aminopeptidase (pepX) gene from Lactobacillus helveticus

The ORF YNL274c (GOR1) codes for glyoxylate reductase in Saccharomyces cerevisiae

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