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Transcriptome analysis of a phenol-producing Pseudomonas putida S12 construct: Genetic and physiological basis for improved production

Wierckx, N.J.P. (author), Ballerstedt, H. (author), de Bont, J.A.M. (author), de Winde, J.H. (author), Ruijssenaars, H.J. (author), Wery, J. (author), TNO Kwaliteit van Leven (author)

The unknown genetic basis for improved phenol production by a recombinant Pseudomonas putida S12 derivative bearing the tpl (tyrosine-phenol lyase) gene was investigated via comparative transcriptomics, nucleotide sequence analysis, and targeted gene disruption. We show upregulation of tyrosine biosynthetic genes and possibly decreased...

article 2008

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A framework to identify physiological responses in microarray-based gene expression studies: Selection and interpretation of biologically relevant genes

Rodenburg, W. (author), Heidema, A.G. (author), Boer, J.M.A. (author), Bovee-Oudenhoven, I.M.J. (author), Feskens, E.J.M. (author), Mariman, E.C.M. (author), Keijer, J. (author), TNO Kwaliteit van Leven (author)

In whole genome microarray studies major gene expression changes are easily identified, but it is a challenge to capture small, but biologically important, changes. Pathway-based programs can capture small effects but may have the disadvantage of being restricted to functionally annotated genes. A structured approach toward the identification of...

article 2008

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Molecular cloning, functional characterization and expression analysis of a novel monosaccharide transporter gene OsMST6 from rice (Oryza sativa L.)

Wang, Y. (author), Xiao, Y. (author), Zhang, Y. (author), Chai, C. (author), Wei, G. (author), Wei, X. (author), Xu, H. (author), Wang, M. (author), Ouwerkerk, P.B.F. (author), Zhu, Z. (author), TNO Kwaliteit van Leven (author)

Monosaccharides transporters play important roles in assimilate supply for sink tissue development. In this study, a new monosaccharide transporter gene OsMST6 was identified from rice (Oryza sativa L.). The predicted OsMST6 protein shows typical features of sugar transporters and shares 79.6% identity with the rice monosaccharide transporter...

article 2008

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Transcriptional activity around bacterial cell death reveals molecular biomarkers for cell viability

TNO Kwaliteit van Leven (author), Kort, R. (author), Keijser, B.J. (author), Caspers, M.P.M. (author), Schuren, F.H. (author), Montijn, R. (author)

Background: In bacteriology, the ability to grow in selective media and to form colonies on nutrient agar plates is routinely used as a retrospective criterion for the detection of living bacteria. However, the utilization of indicators for bacterial viability-such as the presence of specific transcripts or membrane integrity-would overcome bias...

article 2008

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Genomic diversity within the enterobacter cloacae complex

TNO Kwaliteit van Leven (author), Paauw, A. (author), Caspers, M.P.M. (author), Schuren, F.H.J. (author), Leverstein-van Hall, M.A. (author), Delétoile, A. (author), Montijn, R.C. (author), Verhoef, J. (author), Fluit, A.C. (author)

Background: Isolates of the Enterobacter cloacae complex have been increasingly isolated as nosocomial pathogens, but phenotypic identification of the E. cloacae complex is unreliable and irreproducible. Identification of species based on currently available genotyping tools is already superior to phenotypic identification, but the taxonomy of...

article 2008

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Molecular and biochemical characterization of a novel intracellular invertase from Aspergillus niger with transfructosylating activity

Goosen, C. (author), Yuan, X.L. (author), van Munster, J.M. (author), Ram, A.F.J. (author), van der Maarel, M.J.E.C. (author), Dijkhuizen, L. (author), TNO Kwaliteit van Leven (author)

A novel subfamily of putative intracellular invertase enzymes (glycoside hydrolase family 32) has previously been identified in fungal genomes. Here, we report phylogenetic, molecular, and biochemical characteristics of SucB, one of two novel intracellular invertases identified in Aspergillus niger. The sucB gene was expressed in Escherichia...

article 2007

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Lupine allergy: Not simply cross-reactivity with peanut or soy

Peeters, K.A.B.M. (author), Nordlee, J.A. (author), Penninks, A.H. (author), Chen, L. (author), Goodman, R.E. (author), Bruijnzeel-Koomen, C.A.F.M. (author), Hefle, S.L. (author), Taylor, S.L. (author), Knulst, A.C. (author), TNO Kwaliteit van Leven (author)

Background: Reports of lupine allergy are increasing as its use in food products increases. Lupine allergy might be the consequence of cross-reactivity after sensitization to peanut or other legumes or de novo sensitization. Lupine allergens have not been completely characterized. Objectives: We sought to identify allergens associated with...

article 2007

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Dynamics in prophage content of invasive and noninvasive M1 and M28 Streptococcus pyogenes isolates in the Netherlands from 1959 to 1996

Vlaminckx, B.J.M. (author), Schuren, F.H.J. (author), Montijn, R.C. (author), Caspers, M.P.M. (author), Beitsma, M.M. (author), Wannet, W.J.B. (author), Schouls, L.M. (author), Verhoef, J. (author), Jansen, W.T.M. (author), TNO Kwaliteit van Leven (author)

Invasive group A streptococcal (GAS) disease reemerged in The Netherlands in the late 1980s. To seek an explanation for this resurgence, the genetic compositions of 22 M1 and 19 M28 GAS strains isolated in The Netherlands between 1960s and the mid-1990s were analyzed by using a mixed-genome DNA microarray. During this four-decade period, M1 and...

article 2007

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Discovering gene expression patterns in time course microarray experiments by ANOVA-SCA

Nueda, M.J. (author), Conesa, A. (author), Westerhuis, J.A. (author), Hoefsloot, H.C.J. (author), Smilde, A.K. (author), Talón, M. (author), Ferrer, A. (author), TNO Kwaliteit van Leven (author)

Motivation: Designed microarray experiments are used to investigate the effects that controlled experimental factors have on gene expression and learn about the transcriptional responses associated with external variables. In these datasets, signals of interest coexist with varying sources of unwanted noise in a framework of (co)relation among...

article 2007

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Analysis of temporal gene expression during Bacillus subtilis spore germination and outgrowth

Keijser, B.J.F. (author), ter Beek, A. (author), Rauwerda, H. (author), Schuren, F. (author), Montijn, R. (author), van der Spek, H. (author), Brul, S. (author), TNO Kwaliteit van Leven (author)

Bacillus subtilis forms dormant spores upon nutrient depletion. Under favorable environmental conditions, the spore breaks its dormancy and resumes growth in a process called spore germination and outgrowth. To elucidate the physiological processes that occur during the transition of the dormant spore to an actively growing vegetative cell, we...

article 2007

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Determination of the relationship between group A streptococcal genome content, M type, and toxic shock syndrome by a mixed genome microarray

Vlaminckx, B.J.M. (author), Schuren, F.H.J. (author), Montijn, R.C. (author), Caspers, M.P.M. (author), Fluit, A.C. (author), Wannet, W.J.B. (author), Schouls, L.M. (author), Verhoef, J. (author), Jansen, W.T.M. (author), TNO Kwaliteit van Leven (author)

Group A streptococci (GAS), or Streptococcus pyogenes, are associated with a remarkable variety of diseases, ranging from superficial infections to life-threatening diseases such as toxic-shock-like syndrome (TSS). GAS strains belonging to M types M1 and M3 are associated with TSS. This study aims to obtain insight into the gene profiles...

article 2007

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Genomotyping of Pseudomonas putida strains using P. putida KT2440-based high-density DNA microarrays: Implications for transcriptomics studies

Ballerstedt, H. (author), Volkers, R.J.M. (author), Mars, A.E. (author), Hallsworth, J.E. (author), Santos, V.A.M.D. (author), Puchalka, J. (author), van Duuren, J. (author), Eggink, G. (author), Timmis, K.N. (author), de Bont, J.A.M. (author), Wery, J. (author), TNO Kwaliteit van Leven (author)

Pseudomonas putida KT2440 is the only fully sequenced P. putida strain. Thus, for transcriptomics and proteomics studies with other P. putida strains, the P. putida KT2440 genomic database serves as standard reference. The utility of KT2440 whole-genome, high-density oligonucleotide microarrays for transcriptomics studies of other Pseudomonas...

article 2007

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Physiological and phylogenetic characterization of a stable benzene-degrading, chlorate-reducing microbial community

Weelink, S.A.B. (author), Tan, N.C.G. (author), ten Broeke, H. (author), van Doesburg, W. (author), Langenhoff, A.A.M. (author), Gerritse, J. (author), Stams, A.J.M. (author), TNO Bouw en Ondergrond (author)

A stable anoxic enrichment culture was obtained that degraded benzene with chlorate as an electron acceptor. The benzene degradation rate was 1.65 mM benzene per day, which is similar to reported aerobic benzene degradation rates but 20-1650 times higher than reported for anaerobic benzene degradation. Denaturing gradient gel electrophoresis of...

article 2007

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Transcriptomics analysis of interactive effects of benzene, trichloroethylene and methyl mercury within binary and ternary mixtures on the liver and kidney following subchronic exposure in the rat

Hendriksen, P.J.M. (author), Freidig, A.P. (author), Jonker, D. (author), Thissen, U. (author), Bogaards, J.J.P. (author), Mumtaz, M.M. (author), Groten, J.P. (author), Stierum, R.H. (author), TNO Kwaliteit van Leven (author)

The present research aimed to study the interaction of three chemicals, methyl mercury, benzene and trichloroethylene, on mRNA expression alterations in rat liver and kidney measured by microarray analysis. These compounds were selected based on presumed different modes of action. The chemicals were administered daily for 14 days at the Lowest...

article 2007

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Pathway and single gene analyses of inhibited Caco-2 differentiation by ascorbate-stabilized quercetin suggest enhancement of cellular processes associated with development of colon cancer

Dihal, A.A. (author), Tilburgs, C. (author), van Erk, M.J. (author), Rietjens, I.M.C.M. (author), Woutersen, R.A. (author), Stierum, R.H. (author), TNO Kwaliteit van Leven (author)

The aim was to investigate mechanisms contributing to quercetin's previously described effects on cell-proliferation and -differentiation, which contradicted its proposed anticarcinogenic potency. In a 10-day experiment, 40 μM quercetin stabilized by 1 mM ascorbate reduced Caco-2 differentiation up to 50% (p < 0.001). Caco-2 RNA from days 5 and...

article 2007

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Wnt but not BMP signaling is involved in the inhibitory action of sclerostin on BMP-stimulated bone formation

van Bezooijen, R.L. (author), Svensson, J.P. (author), Eefting, D. (author), Visser, A. (author), van der Horst, G. (author), Karperien, M. (author), Quax, P.H.A. (author), Vrieling, H. (author), Papapoulos, S.E. (author), ten Dijke, P. (author), Löwik, W.G.M.C. (author), TNO Kwaliteit van Leven (author)

Sclerostin is an osteocyte-derived negative regulator of bone formation. It inhibits BMP-stimulated bone formation both in vitro and in vivo but has no direct effect on BMP signaling. Instead, sclerostin inhibits Wnt signaling that is required for BMP-stimulated osteoblastic differentiation. Introduction: Sclerostin is a member of the Dan family...

article 2007

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A sandwich-cultured rat hepatocyte system with increased metabolic competence evaluated by gene expression profiling

Kienhuis, A.S. (author), Wortelboer, H.M. (author), Maas, W.J. (author), van Herwijnen, M. (author), Kleinjans, J.C.S. (author), van Delft, J.H.M. (author), Stierum, R.H. (author), TNO Kwaliteit van Leven (author)

A rapid decline of cytochrome P450 (CYP450) enzyme activities remains a drawback of rat hepatocyte-based in vitro cultures. Consequently, judgment of the toxic potential of compounds that need bioactivation by CYP450s may not be adequate using this model. In the present study, an improved hepatocyte-based in vitro system was developed with...

article 2007

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Identification of glucose-fermenting bacteria present in an in vitro model of the human intestine by RNA-stable isotope probing

Egert, M. (author), de Graaf, A.A. (author), Maathuis, A. (author), de Waard, P. (author), Plugge, C.M. (author), Smidt, H. (author), Deutz, N.E.P. (author), Dijkema, C. (author), de Vos, W.M. (author), Venema, K. (author), TNO Kwaliteit van Leven (author)

16S rRNA-based stable isotope probing (SIP) and nuclear magnetic resonance (NMR) spectroscopy-based metabolic profiling were used to identify bacteria fermenting glucose under conditions simulating the human intestine. The TIM-2 in vitro model of the human intestine was inoculated with a GI tract microbiota resembling that of the small intestine...

article 2007

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Comparison of coumarin-induced toxicity between sandwich-cultured primary rat hepatocytes and rats in vivo: A toxicogenomics approach

Kienhuis, A.S. (author), Wortelboer, H.M. (author), Hoflack, J.C. (author), Moonen, E.J. (author), Kleinjans, J.C.S. (author), van Ommen, B. (author), van Delft, J.H.M. (author), Stierum, R.H. (author), TNO Kwaliteit van Leven (author)

Sandwich-cultured primary rat hepatocytes are often used as an in vitro model in toxicology and pharmacology. However, loss of liver-specific functions, in particular, the decline of cytochrome P450 (P450) enzyme activity, limits the value of this model for prediction of in vivo toxicity. In this study, we investigated whether a hepatic in vitro...

article 2006

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Multivariate analysis of microarray data by principal component discriminant analysis: Prioritizing relevant transcripts linked to the degradation of different carbohydrates in Pseudomonas putida S12

van der Werf, M.J. (author), Pieterse, B. (author), van Luijk, N. (author), Schuren, F. (author), van der Werff van der - Vat, B. (author), Overkamp, K. (author), Jellema, R.H. (author), TNO Kwaliteit van Leven (author)

The value of the multivariate data analysis tools principal component analysis (PCA) and principal component discriminant analysis (PCDA) for prioritizing leads generated by microarrays was evaluated. To this end, Pseudomonas putida S12 was grown in independent triplicate fermentations on four different carbon sources, i.e. fructose, glucose,...

article 2006