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See detailCloning of permease structural gene in yeast
Jauniaux, J. C.; Vandenbol, Micheline ULg; Vissers, S. et al

Poster (1984)

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See detailCloning of the genome of Alcelaphine herpesvirus 1 as an infectious and pathogenic bacterial artificial chromosome
Dewals, Benjamin G ULg; Boudry, Christel; Gillet, Laurent ULg et al

Poster (2006)

Alcelaphine herpesvirus 1 (AlHV-1), carried asymptomatically by wildebeest, causes malignant catarrhal fever (MCF) following cross-species transmission to a variety of susceptible species of the order ... [more ▼]

Alcelaphine herpesvirus 1 (AlHV-1), carried asymptomatically by wildebeest, causes malignant catarrhal fever (MCF) following cross-species transmission to a variety of susceptible species of the order Artiodactyla. The study of MCF pathogenesis has been impeded by an inability to produce recombinant virus, mainly due to the fact that AlHV-1 becomes attenuated during passage in culture. In this study, these difficulties were overcome by cloning the entire AlHV-1 genome as a stable, infectious and pathogenic bacterial artificial chromosome (BAC). A modified loxP-flanked BAC cassette was inserted in one of the two large non-coding regions of the AlHV-1 genome. This insertion allowed the production of an AlHV-1 BAC clone stably maintained in bacteria and able to regenerate virions when transfected into permissive cells. The loxP-flanked BAC cassette was excised from the genome of reconstituted virions by growing them in permissive cells stably expressing Cre recombinase. Importantly, BAC-derived AlHV-1 virions replicated comparably to the virulent (low-passage) AlHV-1 parental strain and induced MCF in rabbits that was indistinguishable from that of the virulent parental strain. The availability of the AlHV-1 BAC is an important advance for the study of MCF that will allow the identification of viral genes involved in MCF pathogenesis, as well as the production of attenuated recombinant candidate vaccines. [less ▲]

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See detailCloning of the genome of Alcelaphine herpesvirus 1 as an infectious and pathogenic bacterial artificial chromosome.
Dewals, Benjamin G ULg; Boudry, Christel ULg; Gillet, Laurent ULg et al

in Journal of General Virology (The) (2006), 87(Pt 3), 509-17

Alcelaphine herpesvirus 1 (AlHV-1), carried asymptomatically by wildebeest, causes malignant catarrhal fever (MCF) following cross-species transmission to a variety of susceptible species of the order ... [more ▼]

Alcelaphine herpesvirus 1 (AlHV-1), carried asymptomatically by wildebeest, causes malignant catarrhal fever (MCF) following cross-species transmission to a variety of susceptible species of the order Artiodactyla. The study of MCF pathogenesis has been impeded by an inability to produce recombinant virus, mainly due to the fact that AlHV-1 becomes attenuated during passage in culture. In this study, these difficulties were overcome by cloning the entire AlHV-1 genome as a stable, infectious and pathogenic bacterial artificial chromosome (BAC). A modified loxP-flanked BAC cassette was inserted in one of the two large non-coding regions of the AlHV-1 genome. This insertion allowed the production of an AlHV-1 BAC clone stably maintained in bacteria and able to regenerate virions when transfected into permissive cells. The loxP-flanked BAC cassette was excised from the genome of reconstituted virions by growing them in permissive cells stably expressing Cre recombinase. Importantly, BAC-derived AlHV-1 virions replicated comparably to the virulent (low-passage) AlHV-1 parental strain and induced MCF in rabbits that was indistinguishable from that of the virulent parental strain. The availability of the AlHV-1 BAC is an important advance for the study of MCF that will allow the identification of viral genes involved in MCF pathogenesis, as well as the production of attenuated recombinant candidate vaccines. [less ▲]

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See detailCloning of the koi herpesvirus genome as an infectious bacterial artificial chromosome demonstrates that disruption of the thymidine kinase locus induces partial attenuation in Cyprinus carpio koi.
Costes, Bérénice ULg; Fournier, Guillaume ULg; Michel, Benjamin ULg et al

in Journal of Virology (2008), 82(10), 4955-4964

Koi herpesvirus (KHV) is the causative agent of a lethal disease in koi and common carp. In the present study, we describe the cloning of the KHV genome as a stable and infectious bacterial artificial ... [more ▼]

Koi herpesvirus (KHV) is the causative agent of a lethal disease in koi and common carp. In the present study, we describe the cloning of the KHV genome as a stable and infectious bacterial artificial chromosome (BAC) clone that can be used to produce KHV recombinant strains. This goal was achieved by the insertion of a loxP-flanked BAC cassette into the thymidine kinase (TK) locus. This insertion led to a BAC plasmid that was stably maintained in bacteria and was able to regenerate virions when permissive cells were transfected with the plasmid. Reconstituted virions free of the BAC cassette but carrying a disrupted TK locus (the FL BAC-excised strain) were produced by the transfection of Cre recombinase-expressing cells with the BAC. Similarly, virions with a wild-type revertant TK sequence (the FL BAC revertant strain) were produced by the cotransfection of cells with the BAC and a DNA fragment encoding the wild-type TK sequence. Reconstituted recombinant viruses were compared to the wild-type parental virus in vitro and in vivo. The FL BAC revertant strain and the FL BAC-excised strain replicated comparably to the parental FL strain. The FL BAC revertant strain induced KHV infection in koi carp that was indistinguishable from that induced by the parental strain, while the FL BAC-excised strain exhibited a partially attenuated phenotype. Finally, the usefulness of the KHV BAC for recombination studies was demonstrated by the production of an ORF16-deleted strain by using prokaryotic recombination technology. The availability of the KHV BAC is an important advance that will allow the study of viral genes involved in KHV pathogenesis, as well as the production of attenuated recombinant candidate vaccines. [less ▲]

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See detailCloning of the nitrogen permease regulator gene NPR1 of Saccharomyces cerevisiae.
Vandenbol, Micheline ULg; Jauniaux, J. C.; Vissers, S. et al

in Archives Internationales de Physiologie et de Biochimie (1985), 93

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See detailCloning of the prepro C-RFa gene and brain localization of the active peptide in Salmo salar
Montefusco-Siegmund, R. A.; Romero, A.; Kausel, G. et al

in Cell & Tissue Research (2006), 325(2), 277-285

In all vertebrates, the synthesis and release of prolactin (Prl) from pituitary lactotroph cells is tightly controlled by hypothalamic factors. We have cloned and characterized a hypothalamic cDNA from ... [more ▼]

In all vertebrates, the synthesis and release of prolactin (Prl) from pituitary lactotroph cells is tightly controlled by hypothalamic factors. We have cloned and characterized a hypothalamic cDNA from Atlantic salmon (Salmo salar) encoding C-RFa, a peptide structurally related to mammalian Prl-releasing peptide (PrRP). The deduced preprohormone precursor is composed of 155 amino acid residues presenting a 87.1% similarity to chum salmon C-RFa and a 100% similarity to all fish C-RFa in the bioactive precursor motifs. C-RFa-immunoreactive perikarya and fibres were located in the brain of S. salar, especially in the hypothalamus, olfactory tract, optic tectum and cerebellum. In contrast, immunolabelled fibres were not observed in the pituitary stalk or in the hypophysis. However, interestingly, we detected immunolabelled cells in the rostral pars distalis of the pituitary in the basolateral region in which Prl is synthesized. These results were confirmed by obtaining a strong signal by using reverse transcription/polymerase chain reaction (RTPCR) on mRNA from both hypothalamus and pituitary. These data show, for the first time, by immunohistochemistry and RT-PCR, that C-RFa is produced in pituitary cells. Finally, based on these results, a possible function for CRFa as a locally produced PrRP in this teleost is discussed. [less ▲]

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See detailCloning of the Rat Brain Cdna Encoding for the Slc-1 G Protein-Coupled Receptor Reveals the Presence of an Intron in the Gene
Lakaye, Bernard ULg; Minet, Arlette ULg; Zorzi, Willy ULg et al

in Biochimica et Biophysica Acta (1998), 1401(2), 216-20

In order to isolate new G protein-coupled receptors expressed in the cerebral cortex, a set of degenerate oligonucleotides corresponding to the third and seventh transmembrane segment were synthetized ... [more ▼]

In order to isolate new G protein-coupled receptors expressed in the cerebral cortex, a set of degenerate oligonucleotides corresponding to the third and seventh transmembrane segment were synthetized. Their use in PCR on rat brain cortex mRNA amplified several cDNA fragments. One of them, a 526 bp sequence, encoded for what was at that time an unknown G protein-coupled receptor. An oligonucleotide derived from the sequence was then used as a probe to isolate the receptor cDNA from a rat brain cDNA library. It encodes for a 353aa protein with seven transmembrane segments, three consensus N-glycosylation sites at the amino terminus and several potential phosphorylation sites in the intracellular loops. This protein shares 91% overall identity with a recently cloned human somatostatin-like receptor of 402aa named SLC-1. This suggests that we have cloned the rat orthologue of the human SLC-1. However, the extracellular N-terminus of the human receptor is 49 amino acids longer and shows 50% identity with the rat one. Because the human sequence was deduced from genomic DNA, we suspected the presence of an intron in the gene. This was confirmed by PCR using primers spanning the intron. On the basis of the sequence of a 128 kb fragment of chromosome 22 encompassing the SLC-1 gene, we were able to deduce a corrected amino acids sequence for the human receptor. So both rat and human SLC-1 receptors are 353aa long, with three consensus N-glycosylation sites. They share 96% identity at the amino acid level and are encoded by a gene containing one intron in the coding sequence. [less ▲]

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See detailCloning of the Saccharomyces cerevisiae proline permease structural gene PUR4 and ammonia-regulation in its expression
Jauniaux, J. C.; Vandenbol, Micheline ULg; Vissers, S. et al

in Archives Internationales de Physiologie et de Biochimie (1986), 94

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See detailCloning, biochemical and structural studies of an alcohol dehydrogenase from the Antarctic bacterium Moraxella sp. TAE 123
Tsigos, I.; Georlette, Daphné; Papanikolau, Y. et al

Poster (2002)

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See detailCloning, Nucleotide Sequence and Amplified Expression of the Gene Encoding the Extracellular Metallo (Zn) Dd-Peptidase of Streptomyces Albus G
Duez, Colette ULg; Lakaye, Bernard ULg; Houba, Simone et al

in FEMS Microbiology Letters (1990), 71(1-2), 215-219

The gene encoding the extracellular metallo (Zn) DD-peptidase of Streptomyces albus G has been cloned in Escherichia coli DH5 alpha MCR via pBR322 or 325, and then transferred into Streptomyces lividans ... [more ▼]

The gene encoding the extracellular metallo (Zn) DD-peptidase of Streptomyces albus G has been cloned in Escherichia coli DH5 alpha MCR via pBR322 or 325, and then transferred into Streptomyces lividans TK24 via pIJ486, with substantial amplification of the expressed DD-peptidase. The gene has the information for the synthesis of a 255 amino acid precursor, the amino terminal region of which has the characteristic features of a signal peptide. The primary structure as deduced from nucleotide sequencing confirms that previously determined by chemical methods except for the occurrence of an Asp instead of Asn at position 1 and an additional Ala immediately downstream of Pro67. [less ▲]

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See detailCloning, Sequence and Structural Features of a Lipase from the Antarctic Facultative Psychrophile Psychrobacter Immobilis B10
Arpigny, J. L.; Feller, Georges ULg; Gerday, Charles ULg

in Biochimica et Biophysica Acta (1993), 1171(3), 331-3

A lipase gene (lip1) from the facultative psychrophilic strain Psychrobacter immobilis B10 has been cloned and sequenced. The deduced preprotein sequence is composed of 317 amino acids with a predicted M ... [more ▼]

A lipase gene (lip1) from the facultative psychrophilic strain Psychrobacter immobilis B10 has been cloned and sequenced. The deduced preprotein sequence is composed of 317 amino acids with a predicted M(r) of 35,288. A primary structure alignment of lipases including lip1 shows conserved elements for which a structural role is proposed in the light of recent crystallographic studies. The analysis of the psychrophilic enzyme sequence suggests characteristics in relation with the adaptation to cold. [less ▲]

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See detailCloning, Sequencing and Characterization of PIT1 Gene in Ovis aries
Bastos, Estella; Santos, Ingrid; Parmentier Isabelle et al

in Genetica (2006), 126

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See detailCloning, sequencing and overexpression in Escherichia coli of the alginate lyase-encoding aly gene of Pseudomonas alginovora: identification of three classes of alginate lyases
Chavagnat, Frederic; Duez, Colette ULg; Guinand, Micheline et al

in Biochemical Journal (1996), 319(Pt 2), 575-583

A gene of Pseudomonas alginovora, called aly, has been cloned in Escherichia coli using a battery of PCR techniques and sequenced. It encodes a 210-amino-acid alginate lyase (EC 4.2.2.3), Aly, in the form ... [more ▼]

A gene of Pseudomonas alginovora, called aly, has been cloned in Escherichia coli using a battery of PCR techniques and sequenced. It encodes a 210-amino-acid alginate lyase (EC 4.2.2.3), Aly, in the form of a 233-amino-acid precursor. P. alginovora Aly has been overproduced in E. coli with a His-tag sequence fused at the C-terminal end under conditions in which the formation of inclusion bodies is avoided. His-tagged P. alginovora Aly has the same enzymic properties as the wild-type enzyme and has the specificity of a mannuronate lyase. It can be purified in a one-step procedure by affinity chromatography on Ni(2+)-nitriloacetate resin. The yield is of 5 mg of enzyme per litre of culture. The amplification factor is 12.5 compared with the level of production by wild-type P. alginovora. The six alginate lyases of known primary structure fall into three distinct classes, one of which comprises the pair P. alginovora Aly and Klebsiella pneumoniae Aly. [less ▲]

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See detailClONO2 total vertical column abundances above the Jungfraujoch Station, 1986-1994: Long-term trend and winter-spring enhancements
Rinsland, C. P.; Zander, Rodolphe ULg; Demoulin, Philippe ULg et al

in Journal of Geophysical Research (1996), 101(D2), 3891-3899

Total vertical column abundances of chlorine nitrate (ClONO2) have been retrieved from 0.006 cm(-1) resolution solar absorption spectra recorded at the International Scientific Station of the Jungfraujoch ... [more ▼]

Total vertical column abundances of chlorine nitrate (ClONO2) have been retrieved from 0.006 cm(-1) resolution solar absorption spectra recorded at the International Scientific Station of the Jungfraujoch (ISSJ) in the Swiss Alps (altitude 3.58 km, latitude 46.5 degrees N, longitude 8.0 degrees E) on 105 days between June 1986 and November 1994. The analysis is based on spectral fittings of the ClONO2 nu(4) band Q branch at 780.21 cm(-1) and the interferences occurring in the same spectral region. The ISSJ measurements show a regular long-term increase in the ClONO2 column with an occasional factor of 2 to 3 enhancements during the midwinter to early spring. Excluding data from this time of the year, the ISSJ database reflects a linear rate of increase and la uncertainty equal to (4.0 +/- 0.7)% yr(-1) referenced to 1990.0. The corresponding ClONO2 total vertical columns for mid-1986 and mid-1994 are equal to 0.92 and 1.26 x 10(15) molecules cm(-2), respectively. The high ClONO2 columns and high HF/HCl column ratios sometimes measured during winter indicate the occasional presence of chemically processed air above the station. This is corroborated by trajectories calculated for the stratospheric air masses sounded on these occasions. [less ▲]

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See detailClosantel Intoxication in a Dog
Mc Entee, Kathleen ULg; Grauwels, Magda ULg; Clercx, Cécile ULg et al

in Veterinary and Human Toxicology (1995), 37(3), 234-6

A case of overdosage with closantel, a salicynalide derivative, in a dog is described. The dog received 6 times the recommended dosage. Closantel induced optic neuritis, retinal degeneration, partial ... [more ▼]

A case of overdosage with closantel, a salicynalide derivative, in a dog is described. The dog received 6 times the recommended dosage. Closantel induced optic neuritis, retinal degeneration, partial deafness, hepatotoxicosis and myopathy. Only the blindness was irreversible. The therapy included albumin administration to reduce the acute toxicity of closantel. [less ▲]

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See detailA close look at the RGS spectra of the O4Ief star Zeta Pup
Rauw, Grégor ULg; Flores, A.; Nazé, Yaël ULg et al

Poster (2010)

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