942 resultados para Rescue 1122
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1914/01 (N1).
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Contient : Dénombrement des fiefs de Marie de Soms, dame de Rogy ; Charte de Hugues de Die confirmant les coutumes du comté de Brienne ; Préceptes de Charlemagne pour les Espagnols fugitifs (Böhmer-Mühlbacher, n° 456), et de Louis le Pieux pour Saint-Seurin de Bordeaux ; Charte du roi Jean pour G. Allegrin, chevalier du guet du Lendit ; Hommage prêté à Philippe le Bel par Jean, comte de Hainant, pour la terre d'Ostrevant ; Vente à Philippe-le-Bel par Renaud, comte de Gueldre, de ses droits sur Harfleur, etc ; Bulle de Clément VII pour le monastère d'Essomes ; Chartes de Notre-Dame d'Oulchy ; Accord relatif à la seigneurie de Chatenay et de Vitry ; Notes sur la maison de Roye ; Pièces relatives à la famille de Villeprouvée ; Charte de Raoul, comte de Soissons, pour les habitants de Chacrise ; « Prose » latine satirique sur la journée des Dupes ; Extraits des chartes de l'église de Limoges ; Extrait de la chronique de Géraud de Frachet ; Extraits des chartes de Saint-Loup de Troyes ; Actes du couronnement de saint Louis ; Charte de Louis VIII réglant les droits de ses enfants à sa succession ; Extraits d'un martyrologe de Brion en Anjou ; Extraits d'une Chronique de Sicile ; Extraits de Mémoires sur la famille de Beauvilliers, par Me Robert Cousin ; Extraits de la Chronique d'Aimeri de Peyrat ; Extraits de l'Historia Francorum Floriacensis ; Pièces sur la domination de Simon de Montfort dans le Midi ; Épigrammes latines de Robert Gaguin ; Famille Isoré ; Fragment relatif à la maison de Roucy ; Pièces relatives à la succession à l'Empire en 1257 ; Extrait de l'histoire de l'abbaye de San Michele della Chiusa, du moine Guillaume ; Extrait de la Chronique de Saint-Trond ; Épitaphe du duc Jean I de Bourbon ; « Origo comitum Nivernensium » ; Chronique des évêques du Mans, des origines au XVIe siècle ; Notice sur le prieuré de Saint-Pierre de Bar-sur-Aube ; Chronique de Jourdain d'Osnabrück ; Extrait de la Chronique de Géraud de Frachet (la suite se trouve au fol. 163) ; Extrait de la Chronique de Gérard d'Auvergne ; Extraits de diverses chroniques ; Extrait de la Chronique de Matthieu d'Escouchy ; Extrait des Chroniques de France et de Bourgogne, de Siméon Le Couvreur ; Extraits des Chroniques de Jean Molinet ; Extraits de la Chronique d'Aubri de Trois-Fontaines ; Histoire des Dauphins de Viennois, par Mathieu Thomassin ; Extrait des Libertates Delphinatus [de Gui Pape], Grenoble [1508], in-4° ; Extraits de la Chronique d'Aubri de Trois-Fontaines ; Ordonnance de Philippe-Auguste sur la juridiction ecclésiastique ; Ordonnances de saint Louis touchant les Juifs ; Notice sur les Soreau, seigneurs de Saint-Géran ; Extrait des archives de Saint-Thomas en Forêt ; Pièces concernant principalement la maison de Beaujeu ; Notes et pièces sur la maison d'Auvergne ; Charte de Philippe, comte de Flandre, pour l'abbaye d'Ourscamp ; Extrait d'une continuation d'André de Marchiennes (cf. Mon. Germ., SS., t. XXVI, p. 212) ; Mémoire sur la succession de Provence au temps du roi René ; Diplômes de Charles le Chauve pour l'abbaye de Beaulieu ; Extraits relatifs à l'Université de Paris ; Notes sur l'abbaye de Redon ; Diplôme de Charles le Chauve pour Saint-Julien de Brioude ; Chartes de Faremoutier ; Extraits du registre XXXI [du Trésor des chartes] ; Extraits du registre III du Trésor des chartes ; Testament de Jeanne de Vierzon ; Testament d'Aymar de Roussillon ; Privilège du concile de Compiègne pour Notre-Dame de Soissons ; Notes sur l'église d'Agde ; Extrait d'une chronique latine ; Serment prêté par les bourgeois de Paris à la reine Blanche ; Harangue de l'Université de Paris au cardinal Antoine Duprat ; Extraits des titres de la léproserie de Melun ; Lettre de Foulques, abbé de Corbie, à Gervais, archevêque de Reims ; Extrait d'un inventaire des titres conservés chez le chancelier de Morvilliers, au temps de Louis XI ; Charte d'Avesgaud, évêque du Mans, pour l'église de La Couture ; Diplômes de l'abbaye d'Aniane ; Diplôme de Louis de Pieux pour Cormeri ; Charte de Henri d'Antigné, seigneur de Sainte-Croix, pour Notre-Dame du Miroir ; Charte de Hugues, comte de Chalon et évêque d'Auxerre, pour Cluny (Chartes de Cluny, n° 2484) ; Charte de Guillaume, duc de Normandie, pour l'église de Saint-Evroul ; Diplômes extraits de l'Histoire d'Italie, de Girolamo Briani ; Extraits du cartulaire de Redon ; Épitaphes de l'abbaye de Longpont ; Épitaphes de Saint-Yved de Braine ; Lettres de Louis XII pour César Borgia ; Testament de Jean de Luxembourg, comte de Conversano ; Diplômes de Louis VI, extraits d'un cartulaire de Saint-Denis ; Accord entre Ferri, duc de Lorraine, et Édouard, comte de Bar ; Autorisation de fortifier Sedan, donnée par Charles VII à Jean de la Marche ; Notes de Dom Anselme le Michel et copies de pièces sur Corbie ; Sauvegarde de Jean d'Albret, gouverneur de Champagne, pour les habitants de Chiny, etc ; Notes sur les seigneurs de Mohon ; Pièces relatives à l'église de La Madeleine-des-deux-Amants ; Vente par Guillaume de Tancarville à Jacques de la Roere de la seigneurie de Chemay en Champagne ; Hommages divers rendus au roi ; Testament de Guiotte de Ligny, châtelaine de Lille ; Charte de Philippe, abbé de La Couture du Mans ; Inventaires de titres relatifs à la maison de Laval ; Pièces relatives à la succession de Hugues le Brun, comte de la Marche ; Notes sur Otton IV ; « Lettre de Louis XII à la Sainte-Chapelle de Dijon » ; Lettres de François I érigeant le comté de Montpensier en duché-pairie ; Pièces concernant Philippe, empereur des Romains, extraites du registre XLIX du Trésor des chartes ; Épitaphe de Marie d'Albret, duc de Nevers ; Fragment d'une chronique latine des rois de France jusqu'à Louis VII ; Extraits de Baudri de Bourgueil ; Extraits des poésies de Florus ; Fragment de capitulaire de Charlemagne (Boretius-Krause, t. I, p. 81) ; Extraits du cartulaire du Saint-Sépulcre de Jérusalem ; Fragment d'une enquête sur les droits de l'évêque de Beauvais ; Lettres d'Oliba, évêque de Vich ; Extraits de titres concernant Saint-Mesmin de Micy ; Charte de Renier, évêque d'Orléans, pour l'église de Saint-Marcel ; Épitaphe du comte Eccard ; Prise de possession par Marguerite de Chauvigny, comtesse de Penthièvre, de la seigneurie de Saint-Chartier ; Concile romain de 1112 ; Extrait des Annales de Saint-Orens d'Auch ; Lettre de Laurent, doyen de Poitiers, sur la mort de l'évêque Gilbert ; Mandement de Charles V pour faire fortifier Saint-Mesme près Chinon ; Lettre des évêques de la province de Sens aux cardinaux ; Notice sur l'histoire de l'abbaye de Fleury au temps de Louis VII ; Charte de Sigerius pour le monastère de Notre-Dame (Bréquigny, I, p. 441) ; Extrait d'un traité de Josse Clichtoue ; Extrait d'un inventaire du Trésor des chartes ; Rapport de Chapelain au sujet de l'ouvrage de Camillo Lilii sur les Origines de la nation française ; Extrait de la Chronique d'Hermann de Reichenau ; Fragments extraits de diverses chroniques ; Épitaphe de Bérenger par Hildebert (Migne, Patr. lat., t. 171, col. 1396) ; Sermon d'Henri, évêque d'Albano (Tissier, Bibl. Cisterc., t. III, p. 70) ; Pièces concernant Metz et la Lorraine
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1923/01 (N1A4)-1923/10.
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Bogotá Emprende
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The relative ease to concentrate and purify adenoviruses, their well characterized mid-sized genome, and the ability to delete non-essential regions from their genome to accommodate foreign gene, made adenoviruses a suitable candidate for the construction of vectors. The use of adenoviral vectors in gene therapy, vaccination, and as a general vector system for expressing foreign genes have been documented for some time. In this study, the objective was to rescue a BAV3 E1 or E3 recombinant vector carrying the kanamycin resistant gene, a dominant selectable marker with useful applications in studying vectored gene expression in mammalian cells. To accomplish the objective of this study, more information about BAV3 DNA sequences was required in order to make the manipulation of the virus genome accessible. Therefore, sequencing of the BAV3 genome from 1 1 .7% to 30.8% was carried out. Analysis of the determined sequences revealed the primary structure of important viral gene products coded by E2 including BAV3 DNA pol and precursor to terminal protein. Comparative analysis of these proteins with their counterparts from human and non human adenoviruses revealed important insights as to the evolutionary lineage of BAV3. In order to insert the kanamycin resistance gene in either E1 or E3, it was necessary to delete BAV3 sequences to accommodate the foreign gene so as not to exceed the limit of the packaging capacity of the virus. To construct a recombinant BAV3 in which a foreign gene was inserted in the deleted E1 region, an E1 shuttle vector was constructed. This involved the deletion from the viral sequences a region between 1.3% to 9% and inserting the kanamycin resistance gene to replace the deletion. The E1 shuttle vector contained the left (0%- 53.9%) segment of the genome and was expected to generate BAV3 recombinants that can be grown and propagated in cells that can complement the missing E1 functions. To construct a similar shuttle vector for E3 deletion, DNA sequences extending from 78.9% to 82.5% (1281 bp) were deleted from within the E3 region that had been cloned into a plasmid vector. The deleted region corresponds to those that have been shown to be non-essential for viral replication in cell culture. The resulting plasmid was used to construct another recombinant plasmid with BAV3 DNA sequences extending from 37.1% to 100% and with a deletion of E3 sequences that were replaced by kanamycin resistance gene. This shuttle plasmid was used in cotransfections with digested viral DNA in an attempt to rescue a recombinant BAV3 carrying the kanamycin resistance gene to replace the deleted E3. In spite of repeated attempts of transfection, El or E3 recombinant BAV3 were not isolated. It seems that other approaches should be applied to make a final conclusion on BAV3 infectivity.
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ABSTRACT Recombinant adenoviruses are currently under intense investigation as potential gene delivery and gene expression vectors with applications in human and veterinary medicine. As part of our efforts to develop a bovine adenovirus type 2 (BAV2) based vector system, the nucleotide sequence of BAV2 was determined. Sixty-six open reading frames (ORFs) were found with the potential to encode polypeptides that were at least 50 amino acid (aa) residue long. Thirty-one of the BAV2 polypeptide sequences were found to share homology to already identified adenovirus proteins. The arrangement of the genes revealed that the BAV2 genomic organization closely resembles that of well-characterized human adenoviruses. In the course of this study, continuous propagation of BAV2 over many generations in cell culture resulted in the isolation of a BAV2 spontaneous mutant in which the E3 region was deleted. Restriction enzyme, sequencing and PCR analyses produced concordant results that precisely located the deletion and revealed that its size was exactly 1299 bp. The E3-deleted virus was plaque-purified and further propagated in cell culture. It appeared that the replication of such a virus lacking a portion of the E3 region was not affected, at least in cell culture. Attempts to rescue a recombinant BAV2 virus with the bacterial kanamycin resistance gene in the E3 region yielded a candidate as verified with extensive Southern blotting and PCR analyses. Attempts to purify the recombinant virus were not successful, suggesting that such recombinant BAV2 was helper-dependent. Ten clones containing full-length BAV2 genomes in a pWE15 cosmid vector were constructed. The infectivity of these constructs was tested by using different transfection methods. The BAV2 genomic clones did appear to be infectious only after extended incubation period. This may be due to limitations of various transfection methods tested, or biological differences between virus- and E. co//-derived BAV2 DNA.
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The construction of adenovirus vectors for cloning and foreign gene expression requires packaging cell lines that can complement missing viral functions caused by sequence deletions and/or replacement with foreign DNA sequences. In this study, packaging cell lines were designed to provide in trans the missing bovine adenovirus functions, so that recombinant viruses could be generated. Fetal bovine kidney and lUng cells, acquired at the trimester term from a pregnant cow, were tranfected with both digested wild type BAV2 genomic DNA and pCMV-EI. The plasmid pCMV-EI was specifically constructed to express El of BAV2 under the control of the cytomegalovirus enhancer/promoter (CMV). Selection for "true" transformants by continuous passaging showed no success in isolating immortalised cells, since the cells underwent crisis resulting in complete cell death. Moreover, selection for G418 resistance, using the same cells, also did not result in the isolation of an immortalised cell line and the same culture-collapse event was observed. The lack of success in establishing an immortalised cell line from fetal tissue prompted us to transfect a pre-established cell line. We began by transfecting MDBK (Mardin-Dardy bovine kidney) cells with pCMV-El-neo, which contain the bacterial selectable marker neo gene. A series of MDBK-derived cell lines, that constitutively express bovine adenoviral (BAV) early region 1 (El), were then isolated. Cells selected for resistance to the drug G418 were isolated collectively for full characterisation to assess their suitability as packaging cell lines. Individual colonies were isolated by limiting dilution and further tested for El expression and efficiency of DNA uptake. Two cell lines, L-23 and L-24, out of 48 generated foci tested positive for £1 expression using Northern Blot analysis. DNA uptake studies, using both lipofectamine and calcium phosphate methods, were performed to compare these cells, their parental MDBK cells, 8 and the unrelated human 293 cells as a benchmark. The results revealed that the new MDBKderived clones were no more efficient than MDBK cells in the transient expression of transfected DNA and that they were inferior to 293 cells, when using lacZ as the reporter gene. In view of the inherently poor transfection efficiency of MDBK cells and their derivatives, a number of other bovine cells were investigated for their potential as packaging cells. The cell line CCL40 was chosen for its high efficiency in DNA uptake and subsequently transfected with the plasmid vector pCMV El-neo. By selection with the drug G418, two cell lines were isolated, ProCell 1 and ProCell 2. These cell lines were tested for El expression, permissivity to BAV2 and DNA uptake efficiency, revealing a DNA uptake efficiency of 37 % , comparable to that of CCL40. Attempts to rescue BAV2 mutants carrying the lacZ gene in place of £1 or £3 were carried out by co-transfecting wild type viral DNA with either the plasmid pdlElE-Z (which contains BAV2 sequences from 0% to 40.4% with the lacZ gene in place of the £1 region from 1.1% to 8.25%) or with the plasmid pdlE3-5-Z (which contains BAV2 sequences from 64.8% to 100% with the lacZ gene in place of the E3 region from 75.8% to 81.4%). These cotransfections did not result in the generation of a viral mutant. The lack of mutant generation was thought to be caused by the relative inefficiency ofDNA uptake. Consequently, cosBAV2, a cosmid vector carrying the BAV2 genome, was modified to carry the neo reporter gene in place of the £3 region from 75.8% to 81.4%. The use of a single cosmid vector earring the whole genome would eliminate the need for homologous recombination in order to generate a viral vector. Unfortunately, the transfection of cosBAV2- neo also did not result in the generation of a viral mutant. This may have been caused by the size of the £3 deletion, where excess sequences that are essential to the virus' survival might have been deleted. As an extension to this study, the spontaneous E3 deletion, accidently discovered in our viral stock, could be used as site of foreign gene insertion.
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Helicopter landing on the sunken lawn in front of Memorial Hall, Chapman College, Orange, California.