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Molecular cloning and functional expression in Lactobacillus plantarum 80 of xylT, encoding the D-xylose-H+ symporter of Lactobacillus brevis

Chaillou, S. (author), Bor, Y.-C. (author), Batt, C.A. (author), Postma, P.W. (author), Pouwels, P.H. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

A 3-kb region, located downstream of the Lactobacillus brevis xylA gene (encoding D-xylose isomerase), was cloned in Escherichia coli TG1. The sequence revealed two open reading frames which could code for the D-xylulose kinase gene (xylB) and another gene (xylT) encoding a protein of 457 amino acids with significant similarity to the D-xylose-H...

article 1998

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Binding of human urokinase-type plasminogen activator to its receptor: Residues involved in species specificity and binding

Quax, P.H.A. (author), Grimbergen, J.M. (author), Lansink, M. (author), Bakker, A.H.F. (author), Blatter, M.C. (author), Belin, D. (author), van Hinsbergh, V.W.M. (author), Verheijen, J.H. (author), Gaubius Instituut TNO (author)

Urokinase-type plasminogen activator (UPA), particularly when bound to its receptor (UPAR), is thought to play a major role in local proteolytic processes, thus facilitating cell migration as may occur during angiogenesis, neointima and atherosclerotic plaque formation, and tumor cell invasion. To facilitate understanding of the need and...

article 1998

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Cloning, sequence analysis, and characterization of the genes involved in isoprimeverose metabolism in Lactobacillus pentosus

Chaillou, S. (author), Lokman, B.C. (author), Leer, R.J. (author), Posthuma, C. (author), Postma, P.W. (author), Pouwels, P.H. (author)

Two genes, xylP and xylQ, from the xylose regulon of Lactobacillus pentosus were cloned and sequenced. Together with the repressor gene of the regulon, xylR, the xylPQ genes form an operon which is inducible by xylose and which is transcribed from a promoter located 145 bp upstream of xylP. A putative xylR binding site (xylO) and a cre-like...

article 1998

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Gender-related association between the -93T →Gg/D9N haplotype of the lipoprotein lipase gene and elevated lipid levels in familial combined hyperlipidemia

Hoffer, M.J.V. (author), Bredie, S.J.H. (author), Snieder, H. (author), Reymer, P.W.A. (author), Demacker, P.N.M. (author), Havekes, L.M. (author), Boomsma, D.I. (author), Stalenhoef, A.F.H. (author), Frants, R.R. (author), Kastelein, J.J.P. (author), Gaubius Instituut TNO (author)

Familial combined hyperlipidemia (FCHL) is a frequent cause of premature coronary artery disease. Affected family members are characterized by different combinations of elevated cholesterol and/or triglyceride levels. A reduction in lipoprotein lipase (LPL) activity has been observed in a subgroup of FCHL patients. Recently, we have demonstrated...

article 1998

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Identification and partial characterization of multiple major allergens in peanut proteins

de Jong, E.C. (author), van Zijverden, M. (author), Spanhaak, S. (author), Koppelman, S.J. (author), Pellegrom, H. (author), Penninks, A.H. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

Background: Peanuts are a major cause of food allergies both in children as in adults which can induce an anaphylactic shock. The identification and characterization of peanut allergens could lead to more insight into the mechanism and contribute to the improvement of diagnostic tests and treatment for peanut allergy. Objectives: In the present...

article 1998

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An epitope delivery system for use with recombinant mycobacteria

TNO Preventie en Gezondheid (author), Hetzel, C. (author), Janssen, R. (author), Ely, S.J. (author), Kristensen, N.M. (author), Bunting, K. (author), Cooper, J.B. (author), Lamb, J.R. (author), Young, D.B. (author), Thole, J.E.R. (author)

We have developed a novel epitope delivery system based on the insertion of peptides within a permissive loop of a bacterial superoxide dismutase molecule. This system allowed high-level expression of heterologous peptides in two mycobacterial vaccine strains, Mycobacterium bovis bacille Calmette- Guerin (BCG) and Mycobacterium vaccae. The...

article 1998

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Identification of HLA class II-restricted determinants of Mycobacterium tuberculosis-derived proteins by using HLA-transgenic, class II-deficient mice

TNO Preventie en Gezondheid (author), Geluk, A. (author), Taneja, V. (author), van Meijgaarden, K.E. (author), Zanelli, E. (author), Abou-Zeid, C. (author), Thole, J.E.R. (author), de Vries, R.R.P. (author), David, C.S. (author), Ottenhoff, T.H.M. (author)

T helper 1 cells play a major role in protective immunity against mycobacterial pathogens. Since the antigen (Ag) specificity of CD4+ human T cells is strongly controlled by HLA class II polymorphism, the immunogenic potential of candidate Ags needs to be defined in the context of HLA polymorphism. We have taken advantage of class II-deficient ...

article 1998

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Synthesis of collagen by bovine chondrocytes cultured in alginate; posttranslational modifications and cell-matrix interaction

Beekman, B. (author), Verzijl, N. (author), Bank, R.A. (author), Von Der Mark, K. (author), TeKoppele, J.M. (author), TNO Preventie en Gezondheid (author)

The extracellular matrix synthesized by articular chondrocytes cultured in alginate beads was investigated. Collagen levels increased sigmoidally with time and remained constant after 2 weeks of culture. The presence of cartilage-specific type II collagen was confirmed immunohistochemically. Predominantly type H collagen was present in the...

article 1997

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Glucoamylase gene fusions alleviate limitations for protein production in Aspergillus awamori at the transcriptional ans (post)translational levels

Gouka, R.J. (author), Punt, P.J. (author), van den Hondel, C.A.M.J.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

In this study we have analyzed the effects of a glucoamylase gene fusion on the mRNA levels and protein levels for the human interleukin-6 gene (hil6) and the guar α-galactosidase gene (aglA). Previously it was shown that production of nonfused α-galactosidase and hIL-6 in Aspergillus awamori was limited at transcriptional and (post...

article 1997

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Factor VII polymorphisms in populations with different risks of cardiovascular disease

de Maat, M.P.M. (author), Green, F. (author), de Knijff, P. (author), Jespersen, J. (author), Kluft, C. (author), Gaubius Instituut TNO (author)

Increased plasma factor VII coagulant activity (FVII:C) has been associated with the risk of ischemic heart disease (IHD). Differences in plasma FVII:C among individuals are associated with three common polymorphisms in the FVII gene. Therefore, we investigated FVII polymorphisms in four populations that differ in their risk of developing...

article 1997

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V. Functions of S-layers

Medisch Biologisch Laboratorium TNO (author), Beveridge, T.J. (author), Pouwels, P.H. (author), Sára, M. (author), Kotiranta, A. (author), Lounatmaa, K. (author), Kari, K. (author), Kerosuo, E. (author), Haapasalo, M. (author), Egelseer, E.M. (author), Schocher, I. (author), Sleytr, U.B. (author), Morelli, L. (author), Callegari, M.L. (author), Nomellini, J.F. (author), Bingle, W.H. (author), Smit, J. (author), Leibovitz, E. (author), Lemaire, M. (author), Miras, I. (author), Salamitou, S. (author), Béguin, P. (author), Ohayon, H. (author), Gounon, P. (author), Matuschek, M. (author), Sahm, K. (author), Bahl, H. (author), Grogono-Thomas, R. (author), Dworkin, J. (author), Blaser, M.J. (author), Woodland, R.M. (author), Newell, D.G. (author), Kessel, M. (author), Koval, S.F. (author)

Although S-layers are being increasingly identified on Bacteria and Archaea, it is enigmatic that in most cases S-layer function continues to elude us. In a few instances, S-layers have been shown to be virulence factors on pathogens (e.g. Campylobacter fetus ssp. fetus and Aeromonas salmonicida), protective against Bdellovibrio, a depository...

article 1997

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IV. Molecular biology of S-layers

Centraal Instituut voor Voedingsonderzoek TNO (author), Bahl, H. (author), Scholz, H. (author), Bayan, N. (author), Chami, M. (author), Leblon, G. (author), Gulik-Krzywicki, T. (author), Shechter, E. (author), Fouet, A. (author), Mesnage, S. (author), Tosi-Couture, E. (author), Gounon, P. (author), Mock, M. (author), de Macario, E.C. (author), Macario, A.J.L. (author), Fernández-Herrero, L.A. (author), Olabarría, G. (author), Berenguer, J. (author), Blaser, M.J. (author), Kuen, B. (author), Lubitz, W. (author), Sára, M. (author), Pouwels, P.H. (author), Kolen, C.P.A.M. (author), Boot, H.J. (author), Palva, A. (author), Truppe, M. (author), Howorka, S. (author), Schroll, G. (author), Lechleitner, S. (author), Resch, S. (author)

In this chapter we report on the molecular biology of crystalline surface layers of different bacterial groups. The limited information indicates that there are many variations on a common theme. Sequence variety, antigenic diversity, gene expression, rearrangements, influence of environmental factors and applied aspects are addressed. There is...

article 1997

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Isolation and molecular characterisation of the gene encoding eburicol 14α-demethylase (CYP51) from Penicillium italicum

van Nistelrooy, J.G.M. (author), van den Brink, J.M. (author), van Kan, J.A.L. (author), van Gorcom, R.F.M. (author), de Waard, M.A. (author), Prins Maurits Laboratorium TNO (author)

The CYP51 gene encoding eburicol 14α-demethylase (P450(14DM)) was cloned from a genomic library of the filamentous fungal plant pathogen Penicillium italicum, by heterologous hybridisation with the corresponding gene encoding lanosterol 14α-demethylase from the yeast Candida tropicalis. The nucleotide sequence of a 1739-bp genomic fragment and...

article 1996

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Interchange of the active and silent S-layer protein genes of Lactobacillus acidophilus by inversion of the chromosomal sip segment

Boot, H.J. (author), Kolen, C.P.A.M. (author), Pouwels, P.H. (author), Prins Maurits Laboratorium TNO (author)

The most-dominant surface-exposed protein in many bacterial species is the S-protein. This protein crystallises into a regular monolayer on the outside surface of the bacteria: the S-layer. Lactobacillus acidophilus harbours two S-protein-encoding genes, slpA and slpB, only one of which (slpA) is expressed. In this study, we show by polymerase...

article 1996

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Aggregated, conformationally changed fibrinogen exposes the stimulating sites for t-PA-catalysed plasminogen activation

Haddeland, U. (author), Sletten, K. (author), Bennick, A. (author), Nieuwenhuizen, W. (author), Brosstad, F. (author), Gaubius Instituut TNO (author)

The present paper shows that conformationally changed fibrinogen can expose the sites Aα-(148-160) and γ-(312-324) involved in stimulation of the tissue-type plasminogen activator (t-PA)-catalysed plasminogen activation. The exposure of the stimulating sites was determined by ELISA using mABs directed to these sites, and was shown to coincide...

article 1996

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Human T cell repertoire generation in the absence of MHC class II expression results in a circulating CD4+CD8- population with altered physicochemical properties of complementarity-determining region 3

Henwood, J. (author), van Eggermond, M.C.J.A. (author), Boxel van-Dezaire, A.H.H. (author), Schipper, R. (author), den Hoedt, M. (author), Peijnenburg, A. (author), Sanal, Ö. (author), Ersoy, F. (author), Rijkers, G.T. (author), Zegers, B.J.M. (author), Vossen, J.M. (author), van Tol, M.J.D. (author), van den Elsen, P.J. (author), TNO Preventie en Gezondheid (author)

In this study, we have investigated the impact of deficient MHC class II expression on the use of TCRBV6 and TCRBJ gene elements, and on the pattern of amino acid incorporation exhibited in the N1-D-N2 segments of the third complementarity-determining region (CDR3) of these TCRBV6 rearrangements. To this end, we have analyzed circulating T cells...

article 1996

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A convenient and reproducible method to genetically transform bacteria of the genus Bifidobacterium

Argnani, A. (author), Leer, R.J. (author), van Luijk, N. (author), Pouwels, P.H. (author), TNO Voeding (author)

A protocol was developed for the introduction of foreign plasmid DNA into various Bifidobacterium strains. The method, which is applicable to all Bifidobacterium species tested so far, is based on electroporation of bacteria made competent by preincubation in electroporation buffer for several hours at 4°C. Transformation of Bifidobacterium...

article 1996

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Characterization of O-glycosylated precursors of insulin-like growth factor II by matrix-assisted laser desorption/ionization mass spectrometry

Jespersen, S. (author), Koedam, J.A. (author), Hoogerbrugge, C.M. (author), Tjaden, U.R. (author), van der Greef, J. (author), van den Brande, J.L. (author), TNO Voeding (author)

High molecular weight precursors of insulin-like growth factor II (IGF-II) were isolated from Cohn fraction IV of human plasma by ultrafiltration, affinity chromatography and reversed-phase high-performance liquid chromatography. Molecular weight determination by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) of two...

article 1996

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Structure, localization and action of a novel inhibitory neuropeptide involved in the feeding of Lymnaea

Li, K.W. (author), van Minnen, J. (author), van Veelen, P.A. (author), van der Greef, J. (author), Geraerts, W.P.M. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

A neuropeptide that strongly inhibits the spontaneous contractions of the oesophagus in Lymnaea has been characterized as GAPRFVamide. Direct mass spectrometry of nervous tissues and immunocytochemical studies show that the peptide is synthesized by neurones in the buccal ganglia and transported to the oesophagus via the dorso-buccal nerve. In...

article 1996

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Molecular characterization of a saline-soluble lectin from a parasitic fungus: Extensive sequence similarities between fungal lectins

Rosén, S. (author), Kata, M. (author), Persson, Y. (author), Lipniunas, P.H. (author), Wikström, M. (author), van den Hondel, C.A.M.J.J. (author), van den Brink, J.M. (author), Rask, L. (author), Hedén, L.O. (author), Tunlid, A. (author), TNO Voeding (author)

It has been proposed that the interactions between several parasite and pathogenic fungi and their hosts are mediated by soluble lectins present in the fungus. We have cloned and analyzed a gene encoding such a lectin (AOL) from the nematophagous fungus Arthrobotrys oligospora (deuteromycete). The deduced primary structure of the AOL gene...

article 1996