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Métagénomique et approches alternatives pour l'étude fondamentale et l'exploitation de la microflore tellurique / Metagenomics and alternative approaches for the fundamental study of exploitation of soil microbesJacquiod, Samuel 12 November 2012 (has links)
Mes travaux de thèse ont été réalisés dans le cadre du projet Européen METAEXPLORE, visant à découvrir de nouvelles enzymes d’intérêt industriel à partir des communautés microbiennes environnementales via les approches dites de « métagénomique ». Je me suis personnellement focalisé sur la recherche de chitinases dans les communautés bactériennes du sol, en particulier celui de la station expérimentale de Rothamsted (Royaume-Uni). Des approches de séquençage en 454 ont été réalisées afin de caractériser les bactéries dégradant la chitine au travers d’une stratégie d’enrichissement du sol en microcosme. J’ai également pu réaliser des criblages génétiques de banque de clones fosmidiques afin d’identifier des gènes d’intérêt potentiel. J’ai également été impliqué dans le développement d’un nouvel outil biotechnologique appelé « Genefish », dont le but est la capture de séquences d’ADN environnementales d’intérêt sur un plasmide à l’aide d’une souche E. coli ultra-recombinogène. Mes travaux seront présentés sous la forme de trois chapitres, reprenant successivement :- Chapitre 1 : Une synthèse bibliographique du contexte scientifique- Chapitre 2 : La recherche de nouvelles chitinases via des approches métagénomiques- Chapitre 3 : Le développement et l’utilisation de l’outil « Genefish / I realized my PhD in the frame of the European project METAEXPLORE, which aims to discover new enzymes of industrial interest from the environmental microbial communities through the metagenomic approaches. I was personally focused on finding new chitinases within the soil bacterial communities, with a particular emphasis on the Park Grass soil from Rothamsted research station (U.K). Pyrosequencing approaches were applied in order to characterize chitin degrading bacteria through a soil enrichment strategy in microcosm. I was also involved in genetic screening of fosmid clone library for identifying potential genes of interest. Furthermore, I participated to the development of a new biotechnological tool called “Genefish”, which aims to capture environmental DNA sequence of interest into a plasmid thanks an hyper-recombineering E. coli strain. My PhD work will be presented in 3 chapters:- Chapter 1: A literature review of the scientific context- Chapter 2: The search for new chitinases through metagenomic approaches- Chapter 3: The development and the use of the “Genefish” tool
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Physiological and biochemical analysis of modification of Escherichia coli valyl-tRNA synthetase by vs mutants of the bacteriophage T₄Davis, Vicki L January 2011 (has links)
Typescript. / Digitized by Kansas Correctional Industries
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Extraction, purification and characterization of chlorophyllase from alga Phaeodactylum tricornutumKhalyfa, Abdelnaby January 1993 (has links)
No description available.
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On the nature of the enzyme defect(s) in GM1-gangliosidosis types 1, 2 and 3Miller, Jack January 1974 (has links)
No description available.
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Molecular characterization of a pyrophosphate-energized proton pumpSarafian, Vahé January 1992 (has links)
No description available.
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Characterization and adsorption of the cellulase components from Trichoderma reeseiKyriacou, Andreas January 1987 (has links)
No description available.
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Proteolytic enzymes in grass pollen and their relationship to allergenic proteinsSaldanha, Rohit Gregory, Medical Sciences, Faculty of Medicine, UNSW January 2005 (has links)
Pollen grains are ubiquitous triggers of allergic asthma and seasonal rhinitis. Proteolytic enzymes in pollen as well as other sources are capable of disrupting airway epithelial integrity in vivo and in vitro. This provides a plausible mechanism for the initiation of sensitisation of the respiratory immune system to inhaled pollen allergens, comparable to that suggested for Group 1 allergens from house dust mites and cat dander, which are known to possess intrinsic proteolytic activity. This thesis explores the relationship between pollen allergens and proteolytic enzymes. It describes the different strategies used for the characterisation, purification and identification of immunogenic and proteolytic proteins in the complex mixtures of pollen diffusates. The peptidases in the diffusates of Kentucky blue grass, ryegrass and Bermuda grass pollens were characterised by a sensitive fluorescence assay and gelatin zymography. Among these, Bermuda grass pollen demonstrated the presence of a serine peptidase at Mr ~30,000 Da, which corresponded to an intense band by Western blotting using a monoclonal antibody to the timothy grass (Phleum pratense) group 1 allergen, Phl p 1. Purification of this enzyme from Bermuda grass was complicated by the low levels of the enzyme present in the diffusate, as well as by its autohydrolysis. Partial purification of the serine peptidase activity by affinity chromatography using Concanavalin A Sepharose demonstrated that the diffusate contained a trypsin-like peptidase, detected by the fluorescent assay, in addition to the ~30,000 Da serine endopeptidase, detected on gelatin zymography. Proteomic analysis of the ~30,000 Da protein using one- and two-dimensional electrophoresis and mass spectrometry identified it as the major pollen allergen of Bermuda grass, Cyn d 1. The studies reported here provide, for the first time, evidence that a pollen allergen may possess intrinsic proteolytic activity. This activity may play a role in the initiation of airway inflammation and allergic sensitisation.
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The development of multi-component nucleic acid enzymes(MNAzymes)for the detection of analytesMokany, Elisa, Biotechnology & Biomolecular Sciences, Faculty of Science, UNSW January 2007 (has links)
No description available.
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Increasing the thermostability of barley (1->3,1->4)-B-glucanases / Richard John Stewart.Stewart, Richard John January 1999 (has links)
Bibliography: leaves 133-157. / xiii, 157, [22] leaves, [31] leaves of plates : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / The principle aim of the work described in this thesis was to use protein engineering to increase the thermostability of barley (1->3,1->4)-B-glucanases / Thesis (Ph.D.)--University of Adelaide, Dept. of Plant Science, 2000
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The discriminator domain : does it reside at the C-terminus or the N-terminus of Escherichia coli Lon?Miller, Darcey L. 27 August 2001 (has links)
The mechanisms of substrate recognition by regulatory proteases are not well
understood. Presently, two opposing models have arisen to describe E. coil Lon's ability
to discriminate between substrates: one suggests the N-terminus involvement while the
second suggests the C-terminus involvement. In this project, the role of the C-terminal
domain as it relates to the recognition of Lon's normal physiological substrates RcsA, an
activator of colanic acid capsular polysaccharide, and SulA, an inhibitor of cell division,
was addressed. Using site-directed mutagenesis, five mutations in Lon (R537G, E538A,
GS40W, R542G, R542P) were isolated. Their phenotypic impact was either similar in
character to wildtype Lon (R537G, E538A) or ��lon cells (G540W, R542G, R542P). The
stabilization of both RcsA and SulA based on phenotypic assays and immunological
detection of lon* strains (G540W, R542G, R542P) suggests the C-terminal domain may
be involved in substrate degradation as opposed to discriminator activity. / Graduation date: 2002
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