Relevant bibliographies by topics / Synthèss in vitro

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Academic literature on the topic 'Synthèss in vitro'

Author: Grafiati
Published: 4 June 2021
Last updated: 17 February 2022

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Journal articles on the topic "Synthèss in vitro"

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Santosh Kumar, Badampudi, Papammagari Raveendra Reddy, Lakshmana Rao Krishna Rao Ravindranath, Aluru Raghavendra Guru Prasad, and Alvala Mallika. "Synthesis, characterization and in vitro antimicrobial evaluation of sulphonyl urea derivatives as potential inhibitors of beta-ketoacyl-acyl carrier protein synthase III (FabH)." Acta Universitaria 25, no. 1 (March 2015): 12–21. http://dx.doi.org/10.15174/au.2015.658.

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Bellini, Erika, Claudio Varotto, Marco Borsò, Lorenza Rugnini, Laura Bruno, and Luigi Sanità di Toppi. "Eukaryotic and Prokaryotic Phytochelatin Synthases Differ Less in Functional Terms Than Previously Thought: A Comparative Analysis of Marchantia polymorpha and Geitlerinema sp. PCC 7407." Plants 9, no. 7 (July 20, 2020): 914. http://dx.doi.org/10.3390/plants9070914.

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This paper reports functional studies on the enzyme phytochelatin synthase in the liverwort Marchantia polymorpha and the cyanobacterium Geitlerinema sp. strain PCC 7407. In vitro activity assays in control samples (cadmium-untreated) showed that phytochelatin synthase was constitutively expressed in both organisms. In the presence of 100 µM cadmium, in both the liverwort and the cyanobacterium, the enzyme was promptly activated in vitro, and produced phytochelatins up to the oligomer PC4. Likewise, in vivo exposure to 10–36 µM cadmium for 6-120 h induced in both organisms phytochelatin synthesis up to PC4. Furthermore, the glutathione (GSH) levels in M. polymorpha were constitutively low (compared with the average content in higher plants), but increased considerably under cadmium stress. Conversely, the GSH levels in Geitlerinema sp. PCC 7407 were constitutively high, but were halved under metal treatments. At odds with former papers, our results demonstrate that, as in M. polymorpha and other plants, the cyanobacterial phytochelatin synthase exposed to cadmium possesses manifest transpeptidasic activity, being able to synthesize phytochelatins with a degree of oligomerization higher than PC2. Therefore, prokaryotic and eukaryotic phytochelatin synthases differ less in functional terms than previously thought.
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Slayden, R. A., R. E. Lee, J. W. Armour, A. M. Cooper, I. M. Orme, P. J. Brennan, and G. S. Besra. "Antimycobacterial action of thiolactomycin: an inhibitor of fatty acid and mycolic acid synthesis." Antimicrobial Agents and Chemotherapy 40, no. 12 (December 1996): 2813–19. http://dx.doi.org/10.1128/aac.40.12.2813.

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Thiolactomycin (TLM) possesses in vivo antimycobacterial activity against the saprophytic strain Mycobacterium smegmatis mc2155 and the virulent strain M. tuberculosis Erdman, resulting in complete inhibition of growth on solid media at 75 and 25 micrograms/ml, respectively. Use of an in vitro murine macrophage model also demonstrated the killing of viable intracellular M. tuberculosis in a dose-dependent manner. Through the use of in vivo [1,2-14C]acetate labeling of M. smegmatis, TLM was shown to inhibit the synthesis of both fatty acids and mycolic acids. However, synthesis of the shorter-chain alpha'-mycolates of M. smegmatis was not inhibited by TLM, whereas synthesis of the characteristic longer-chain alpha-mycolates and epoxymycolates was almost completely inhibited at 75 micrograms/ml. The use of M. smegmatis cell extracts demonstrated that TLM specifically inhibited the mycobacterial acyl carrier protein-dependent type II fatty acid synthase (FAS-II) but not the multifunctional type I fatty acid synthase (FAS-I). In addition, selective inhibition of long-chain mycolate synthesis by TLM was demonstrated in a dose-response manner in purified, cell wall-containing extracts of M. smegmatis cells. The in vivo and in vitro data and knowledge of the mechanism of TLM resistance in Escherichia coli suggest that two distinct TLM targets exist in mycobacteria, the beta-ketoacyl-acyl carrier protein synthases involved in FAS-II and the elongation steps leading to the synthesis of the alpha-mycolates and oxygenated mycolates. The efficacy of TLM against M. smegmatis and M. tuberculosis provides the prospects of identifying fatty acid and mycolic acid biosynthetic genes and revealing a novel range of chemotherapeutic agents directed against M. tuberculosis.
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Ku, Bosung, Jae-Cheol Jeong, Benjamin N. Mijts, Claudia Schmidt-Dannert, and Jonathan S. Dordick. "Preparation, Characterization, and Optimization of an In Vitro C30 Carotenoid Pathway." Applied and Environmental Microbiology 71, no. 11 (November 2005): 6578–83. http://dx.doi.org/10.1128/aem.71.11.6578-6583.2005.

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ABSTRACT The ispA gene encoding farnesyl pyrophosphate (FPP) synthase from Escherichia coli and the crtM gene encoding 4,4′-diapophytoene (DAP) synthase from Staphylococcus aureus were overexpressed and purified for use in vitro. Steady-state kinetics for FPP synthase and DAP synthase, individually and in sequence, were determined under optimized reaction conditions. For the two-step reaction, the DAP product was unstable in aqueous buffer; however, in situ extraction using an aqueous-organic two-phase system resulted in a 100% conversion of isopentenyl pyrophosphate and dimethylallyl pyrophosphate into DAP. This aqueous-organic two-phase system is the first demonstration of an in vitro carotenoid synthesis pathway performed with in situ extraction, which enables quantitative conversions. This approach, if extended to a wide range of isoprenoid-based pathways, could lead to the synthesis of novel carotenoids and their derivatives.
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Barekzi, Nazir, Swati Joshi, Scott Irwin, Todd Ontl, and Herbert P. Schweizer. "Genetic characterization of pcpS, encoding the multifunctional phosphopantetheinyl transferase of Pseudomonas aeruginosa." Microbiology 150, no. 4 (April 1, 2004): 795–803. http://dx.doi.org/10.1099/mic.0.26823-0.

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Fatty acid synthases (primary metabolism), non-ribosomal peptide synthases and polyketide synthases (secondary metabolism) contain phosphopantetheinyl (Ppant)-dependent carrier proteins that must be made functionally active by transfer of the 4′-Ppant moiety from coenzyme A. These reactions are usually catalysed by dedicated Ppant transferases. Although rich in Ppant-dependent carrier proteins, it was previously shown that Pseudomonas aeruginosa possesses only one Ppant transferase, encoded by pcpS, which functions in both primary and secondary metabolism. Consistent with this notion are our findings that pcpS can genetically complement mutations in the Escherichia coli acpS and entD genes, encoding the apo-acyl carrier protein (ACP) synthase of fatty acid synthesis and a Ppant transferase of enterobactin synthesis, respectively. It also complements a Bacillus subtilis sfp mutation affecting a gene encoding a Ppant transferase essential for surfactin synthesis. A pcpS insertion mutant could only be constructed in a strain carrying the E. coli acpS gene on a chromosomally integrated element in trans, implying that the in vitro essentiality of pcpS is due to its requirement for activation of apo-ACP of fatty acid synthesis. The conditional pcpS mutant is non-fluorescent, does not produce pyoverdine and pyochelin, and does not grow in the presence of iron chelators. The data presented here for the first time confirm that PcpS plays an essential role in both fatty acid and siderophore metabolism.
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Gohil, Dr Neha N., and Dr D. I. Brahmbhatt. "Synthesis, Characterization and In Vitro Antimicrobial Screening of Some PyrazolylPyridyl Substituted Dicoumarins." International Journal of Trend in Scientific Research and Development Volume-2, Issue-4 (June 30, 2018): 1692–705. http://dx.doi.org/10.31142/ijtsrd14420.

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Li, Sun, Zhu, Bian, Wang, and Si. "Identification of New Antifungal Agents Targeting Chitin Synthesis by a Chemical-Genetic Method." Molecules 24, no. 17 (August 29, 2019): 3155. http://dx.doi.org/10.3390/molecules24173155.

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Fungal infection is a leading cause of mortality in immunocompromised population; thus, it is urgent to develop new and safe antifungal agents. Different from human cells, fungi have a cell wall, which is composed mainly of polysaccharide glucan and chitin. The unique cell wall structure is an ideal target for antifungal drugs. In this research, a chemical-genetic method was used to isolate antifungal agents that target chitin synthesis in yeast cells. From a compound library, we isolated two benzothiazole compounds that showed greater toxicity to yeast mutants lacking glucan synthase Fks1 compared to wild-type yeast cells and mutants lacking chitin synthase Chs3. Both of them inhibited the activity of chitin synthase in vitro and reduced chitin level in yeast cells. Besides, these compounds showed clear synergistic antifungal effect with a glucan synthase inhibitors caspofungin. Furthermore, these compounds inhibited the growth of Saccharomyces cerevisiae and opportunistic pathogen Candida albicans. Surprisingly, the genome-wide mass-spectrometry analysis showed decreased protein level of chitin synthases in cells treated with one of these drugs, and this decrease was not a result of downregulation of gene transcription. Therefore, we successfully identified two new antifungal agents that inhibit chitin synthesis using a chemical-genetic method.
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Morii, Hiroyuki, Tadashi Eguchi, and Yosuke Koga. "In Vitro Biosynthesis of Ether-Type Glycolipids in the Methanoarchaeon Methanothermobacter thermautotrophicus." Journal of Bacteriology 189, no. 11 (April 6, 2007): 4053–61. http://dx.doi.org/10.1128/jb.01875-06.

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ABSTRACT The biosynthesis of archaeal ether-type glycolipids was investigated in vitro using Methanothermobacter thermautotrophicus cell-free homogenates. The sole sugar moiety of glycolipids and phosphoglycolipids of the organism is the β-d-glucosyl-(1→6)-d-glucosyl (gentiobiosyl) unit. The enzyme activities of archaeol:UDP-glucose β-glucosyltransferase (monoglucosylarchaeol [MGA] synthase) and MGA:UDP-glucose β-1,6-glucosyltransferase (diglucosylarchaeol [DGA] synthase) were found in the methanoarchaeon. The synthesis of DGA is probably a two-step glucosylation: (i) archaeol + UDP-glucose → MGA + UDP, and (ii) MGA + UDP-glucose → DGA + UDP. Both enzymes required the addition of K+ ions and archaetidylinositol for their activities. DGA synthase was stimulated by 10 mM MgCl2, in contrast to MGA synthase, which did not require Mg2+. It was likely that the activities of MGA synthesis and DGA synthesis were carried out by different proteins because of the Mg2+ requirement and their cellular localization. MGA synthase and DGA synthase can be distinguished in cell extracts greatly enriched for each activity by demonstrating the differing Mg2+ requirements of each enzyme. MGA synthase preferred a lipid substrate with the sn-2,3 stereostructure of the glycerol backbone on which two saturated isoprenoid chains are bound at the sn-2 and sn-3 positions. A lipid substrate with unsaturated isoprenoid chains or sn-1,2-dialkylglycerol configuration exhibited low activity. Tetraether-type caldarchaetidylinositol was also actively glucosylated by the homogenates to form monoglucosyl caldarchaetidylinositol and a small amount of diglucosyl caldarchaetidylinositol. The addition of Mg2+ increased the formation of diglucosyl caldarchaetidylinositol. This suggested that the same enzyme set synthesized the sole sugar moiety of diether-type glycolipids and tetraether-type phosphoglycolipids.
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Matsuo, Shintaro, Satomi Tagawa, Yudai Matsusaki, Yuri Uchi, and Tetsuo Kondo. "Callose-synthesizing enzymes as membrane proteins of Betula protoplasts secrete bundles of β-1,3-glucan hollow fibrils under Ca2+-rich and acidic culture conditions." Holzforschung 74, no. 8 (August 27, 2020): 725–32. http://dx.doi.org/10.1515/hf-2019-0142.

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AbstractPreviously, it was reported that plant protoplasts isolated from Betula platyphylla (white birch) callus secreted bundles of hollow callose fibrils in acidic culture medium containing a high concentration of calcium ions (Ca2+). Here, the callose synthase was characterized from in situ and in vitro perspectives. Localization of callose synthases at the secreting site of callose fiber was indicated from in situ immunostaining observation of protoplasts. For in vitro analyses, membrane proteins were extracted from membrane fraction of protoplasts with a 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) treatment. The CHAPS extract aggregated in the presence of a high concentration of Ca2+, suggesting that Ca2+ may promote the arrangement of callose synthases in the plasma membrane. The callose synthase activity was dependent on pH and Ca2+, similar to the callose synthase of Arabidopsis thaliana. However, the synthesized fibril products were longer than those produced by callose synthases of herbaceous plants. This is the first insight into the specific properties of callose synthases of woody plants that secrete of callose hollow fibers.
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GURCHA, Sudagar S., Alain R. BAULARD, Laurent KREMER, Camille LOCHT, D. Branch MOODY, Walter MUHLECKER, Catherine E. COSTELLO, Dean C. CRICK, Patrick J. BRENNAN, and Gurdyal S. BESRA. "Ppm1, a novel polyprenol monophosphomannose synthase from Mycobacterium tuberculosis." Biochemical Journal 365, no. 2 (July 15, 2002): 441–50. http://dx.doi.org/10.1042/bj20020107.

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Dolichol monophosphomannose (DPM) is an ever-present donor of mannose (Man) in various eukaryotic glycosylation processes. Intriguingly, the related polyprenol monophosphomannose (PPM) is involved in the biosynthesis of lipomannan and lipoarabinomanan, key bacterial factors termed modulins that are found in mycobacteria. Based on similarities to known DPM synthases, we have identified and characterized the PPM synthase of Mycobacterium tuberculosis, now termed Mt-Ppm1. In the present study, we demonstrate that Mt-Ppm1 possesses an unusual two-domain architecture, by which the second domain is sufficient for PPM synthesis. However, when overexpressed separately in mycobacteria, domain 1 of Mt-Ppm1 appears to increase the synthesis of PPM. Interestingly, other mycobacteria such as M. smegmatis, M. avium and M. leprae produce two distinct proteins, which are similar to the two domains found in Mt-Ppm1. Using an in vitro assay, we also demonstrate that Mt-Ppm1 transfers Man from GDP-Man to a structurally diverse range of lipid monophosphate acceptors. The identification of the PPM synthase as a key enzyme in lipoarabinomannan biosynthesis now provides an attractive candidate for gene disruption to generate mutants for subsequent immunological studies. PPM synthase can also be exploited as a target for specific inhibitors of M. tuberculosis.
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Dissertations / Theses on the topic "Synthèss in vitro"

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Ajili, Widad. "Étude des processus de biominéralisation de la nacre chez l'ormeau européen haliotis tuberculata." Thesis, Sorbonne université, 2018. http://www.theses.fr/2018SORUS160.

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La nacre présente dans la coquille de certains mollusques est un matériau fascinant aux propriétés physiques (mécaniques et optiques) remarquable. Ce matériau biologique de nature composite organique et minérale présente un architecture complexe dont la structure et les mécanismes de formation ne sont pas encore parfaitement compris. Dans ces travaux, en prenant comme modèle d’étude comme modèle l’ormeau Européen Haliotis tuberculata, nous avons cherché à contribuer au savoir autour de ce matériau en axant notre étude sur trois aspects en particulier : i) l’étude ultrastructurale de la phase minérale et plus particulièrement des environnements minéraux désordonnés, ii) l’étude de l’interface organominérale dans la nacre adulte en formation ainsi que dans la coquille larvaire et iii) l’étude des premiers biominéraux déposés dans la coquille larvaire aux très jeune stades de développement. Pour réaliser cette étude nous avons eu recours à un couplage de techniques avancées spectroscopiques (Résonnance Magnétique Nucléaire à l’Etat Solide, Microscopie en transmission à balayage de rayons X et Spectroscopie des pertes d'énergie) couplé à des techniques de pointes de microscopie électronique (Microscopie électronique à transmission à haute résolution et Microscope électronique à balayage à effet de champ) et de préparation d’échantillon (Abrasion ionique focalisée) et de biologie moléculaire (Immunohistochimie, Microscopie optique et électronique corrélative)
The mother-of-pearl found in the shell of certain mollusks is a fascinating material with remarkable physical (mechanical and optical) properties. This biological material of organic and mineral composite nature presents a complex architecture whose structure and mechanisms of formation are not yet perfectly understood. In this work, taking as a model study the European abalone Haliotis tuberculata, we sought to contribute to the knowledge around this material by focusing our study on three aspects in particular: i) the ultrastructural study of the phase mineral and more particularly disordered mineral environments, ii) the study of the organo-minerale interface in the adult nacre in formation as well as in the larval shell and iii) the study of the first biominerals deposited in the larval shell at very young stages of development. To carry out this study, we used a coupling of advanced spectroscopic techniques (Solid-State Nuclear Magnetic Resonance, Scanning transmission X-ray microscopy and Electron energy loss spectroscopy) coupled to advanced electron microscopy techniques (High-Resolution Transmission Electronic Microscopies and Field Emission Gun Scanning Electronic Microscopy) and preparation of sample (Focused Ion beam) and molecular biology (immunohistochemistry, Correlative Light and Electronic Microscopy)
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Djerbi, Soraya. "Cellulose synthases in Populus- identification, expression analyses and in vitro synthesis." Doctoral thesis, Stockholm, 2005. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-414.

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Gagnepain, Anne. "Fécondation in vitro : synthèse clinique, biologique, épidémiologique, législative et éthique ; méta-analyse des essais thérapeutiques." Montpellier 1, 1989. http://www.theses.fr/1989MON11258.

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Johnston, Julie Catherine. "In vitro translation of cucumber necrosis virus RNA." Thesis, University of British Columbia, 1989. http://hdl.handle.net/2429/28969.

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The in vitro translation products directed by cucumber necrosis virus (CNV) RNA were analyzed in both rabbit reticulocyte lysate and wheat germ extract cell-free translation systems. In rabbit reticulocyte lysates, one major protein of ca. 33 Mr was produced. In wheat germ extracts, four proteins of ca. 41, 33, 21 and 20 Mr were produced. Hybrid-arrested translation (HART) studies using synthetic CNV antisense RNA corresponding to the entire CNV genome demonstrated that the four major proteins synthesized from CNV virion RNA in wheat germ extracts are virus-specific translation products. The genomic locations of the CNV in vitro translation products were determined using a number of experimental approaches including: (1) HART using antisense RNA corresponding to selected regions of the CNV genome; (2) in vitro translation of synthetic messenger-sense CNV transcripts; (3) immunoprecipitation of in vitro translation products with CNV polyclonal antisera and (4) in vitro translation of size-fractionated CNV virion RNA. Together, these experiments demonstrated that the ca. 33 Mr protein is derived from the 5' proximal coding region, the ca. 41 Mr protein is derived from an internal coding region, and that at least one but probably both of the ca. 20 and 21 Mr proteins are derived from the 3' terminal coding region(s) of the CNV genome. In addition, immunoprecipitation experiments provided further evidence that the ca. 41 Mr protein is the viral coat protein. The size, number, and genomic locations of the CNV in vitro translation products reported here are in agreement with those predicted from nucleotide sequence data (Rochon & Tremaine, 1989). The natural template for the expression of downstream cistrons in the CNV genome was investigated by in vitro translation of sucrose fractionated CNV virion RNA as well as in vitro translation of messenger-sense synthetic transcripts. These studies indicate that in vitro, both subgenomic and genomic-length CNV RNA molecules may act as templates for the synthesis of the ca. 41,21 and 20 Mr proteins as well as the ca. 33 Mr protein.
Land and Food Systems, Faculty of
Graduate
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El, Badaoui Houriya. "Comparaison entre différentes techniques de culture in vitro de Solanum paludosum Moric. Pour la production de solamargine, glycoalcaloi͏̈de principal." Toulouse, INPT, 1993. http://www.theses.fr/1993INPT015A.

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Ce travail porte sur un arbuste tropical: solanum paludosum moric pour la production en glycoalcaloides qui entrent dans l'hemisynthese des hormones steroidiques. Les fruits issus de plantes obtenues par culture in vitro produisent autant de solamargine que les fruits des plantes meres d'origine. Les teneurs sont de 2%. La production des glycoalcaloides par la culture de cals et des cellules desorganisees est faible. Les teneurs sont environ de 0,015%. En revanche la culture de parties aeriennes multiples a permis d'obtenir 0,05% de solamargine alors que les parties aeriennes des plantes entieres ne produisent pas ces glycoalcaloides. La mise au point de la composition des macroelements du milieu de culture in vitro adapte aux besoins de la plante apres analyse minerale a permis d'augmenter 3 fois la production de la biomasse des parties aeriennes multiples et 2 fois la production de la solamargine par ces cultures. La culture des racines in vitro a ete tentee. La biomasse necessaire au dosage de solamargine s'est averee insuffisante ceci confirme les resultats obtenus par d'autres auteurs sur des especes ligneuses comme l'est solanum paludosum
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Marshall, Jonathon J. A. "Regulation of insulin gene expression in rat islets of Langerhans." Thesis, University of Leicester, 1986. http://hdl.handle.net/2381/35103.

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The rate of glucose stimulated in vitro (pro)insulin synthesis in islets of Langerhans isolated from fed male rats was determined. Increases in the glucose concentration of the incubation medium, over the physiological range (2 to 20 mM), stimulated the rates of both (pro)insulin and total protein synthesis during a two hour incubation. This stimulation was preferential for (pro)insulin synthesis and had a sigmoidal dose response curve with a concentration threshold of between 2.5 mM and 5.0 mM and a maximal rate at 20 mM glucose. Inhibition of islet RNA synthesis by.;ctinomycin D depressedthe rate of total protein synthesis within 30 minutes of the application of a 20 mM glucose stimulus. A specific inhibition of (pro)insulin synthesis by actinomycin D, that occured 60 minutes after the application of a 20 mM glucose stimulus, was thought to reflect the inhibition of glucose stimulated preproinsulin mRNA synthesis. The effect of glucose on islet preproinsulin mRNA content during in vitro incubations was also determined. To achieve this a dot blot hybridization assay for preproinsulin mRNA was 3 2 developed which used a P-labeled cloned human insulin gene as the hybridization probe. The assay proved to be sensitive enough to detect preproinsulin mRNA in as few as fifty islets. Incubation of islets, for 2 hours, at 2.5 mM glucose had no effect on islet preproinsulin mRNA content but incubation at 20 mM glucose increased islet preproinsulin mRNA content. Actinomycin D had no effect on this pattern of glucose stimulation and it is suggested that the degradation of preproinsulin mRNA is in some way dependent on RNA synthesis and that its inhibition by glucose plays an important role in the regulation of islet preproinsulin mRNA content.
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Di, Stasio Benoît. "Etude de nouveaux photosensibilisants pour des applications en Thérapie Photodynamique." Thesis, Vandoeuvre-les-Nancy, INPL, 2006. http://www.theses.fr/2006INPL069N/document.

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Les dérivés de porphyrines sont impliqués dans de nombreux processus d'oxydoréduction. Ces composés conduisent à de nombreuses applications, dont la Thérapie Photodynamique (PDT). Il existe deux générations de dérivés de porphyrines qui sont actuellement remplacés par des composés de 3ème génération, plus actifs et entraînant moins d'effets secondaires. Ces composés sont capables de reconnaître spécifiquement et directement (par adressage) ou indirectement (par vectorisation) les cellules cancéreuses. Nous avons orienté notre travail vers la synthèse et l'évaluation biologique de composés tétrapyrroliques associés à des modules tels que des sucres ou des motifs peptidiques de type -RGD- qui permettent une reconnaissance spécifique des cellules cancéreuses, via les lectines ou les intégrines, respectivement. De plus, dans le cadre d'un programme européen Cost-Chemistry intitulé "New molecular systems with therapeutic applications in photodynamic therapy of cancer and microbial infections", nous avons étudié les propriétés photophysiques de nombreux photosensibilisants synthétisés par une équipe roumaine
Derived of porphyrins are tetrapyrrolic macrocycles involved in several redox processes. These compounds are used for different biological applications, like photodynamic therapy (PDT). Many teams in the world seek to synthesize compounds able to directly recognize specifically and (by targeting) or indirectly (by vectorization) cancer cells. These compounds are known as of 3rd generation. We are involved in the synthesis and the biological evaluation of tetrapyrrolic compounds associated to recognition and/or transport agents such as sugars or RGD-like peptide sequences. These moieties allow a specific recognition of the cancerous cells, via the lectins for the sugar moieties and the integrins for the RGD type moieties, respectively. Within the framework of an European Cost-Chemistry program entitled "New mo/ecu/ar systems with therapeutic applications in photodynamic therapy of cancer and microbia/ infections", we also studied the photophysical properties of photosensitizers synthesized by a Rumanian team
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Maake, Takalani Whitney. "Development of an actinobacteria based in vitro transcription and translation systems." University of the Western Cape, 2015. http://hdl.handle.net/11394/4750.

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>Magister Scientiae - MSc
Heterologous metagenomic screening strategies have relied largely on the construction of DNA libraries and screening in Escherichia coli to access novel enzymes. There is an increased demand for the identification of novel lignocellulose degrading enzymes with enhanced biochemical properties which are suitable for applications in industrial processes; biofuels being one of them. The use of heterologous gene expression in function based metagenomic studies has resulted in the discovery of enormous novel bioactive compounds. However, there are limitations associated with using E. coli as a heterologous host which does not allow transcription and translation of all genes in the metagenome. E. coli can only express 40% of the environmental DNA because of promoter recognition, codon usage, and host toxicity of gene products. Therefore alternative strategies for expressing or producing novel enzymes are needed, which can also be employed in metagenomic gene discovery. In vitro protein synthesis is an important tool in molecular biology and used to obtain proteins from genes for functional and expression studies. These systems may hold the key to unlock more of the potential in metagenomic DNA. The broader aim of the study is to develop non- E. coli based cell-free protein synthesis systems to further the metagenomics screening. In this study, Rhodococcus erythropolis H8 was evaluated for its suitability in cell-free expression. Crude extracts containing the macromolecular components (70S or 80S ribosomes, tRNAs, initiation, elongation and termination factors) fromR. erythropolis were prepared using existing crude extract based cell-free protein synthesis (CFPS) protocols. Three genes were selected and used as templates for synthesis: cell11, xp12 and acetyl xylan esterase (axe10), all previously isolated from metagenomic libraries screened inE. coli. As judged by zymograms and enzyme assays, all enzymes were successfully expressedfrom their native promoters and in recombinants clones using the PtipA promoter, and wereactive. Furthermore, the amounts of XP12 protein produced using pFos-XP_12 was 1.2mg/mlfrom E. coli and 1.67mg/ml from R. erythropolis CFPS, showing that the R. erythropolismachinery was more efficient in the expression of XP12 than the E. coli machinery. To the best of our knowledge this is the first demonstration of a cell-free expression using an actinomycete.
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Mould, A. P. "The aggregation properties of type I procollagen in vitro." Thesis, University of Manchester, 1986. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.377670.

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Ahmed, El Sayed Neveen. "Study of RNA synthesis of hepatitis C virus in vitro and in cells of hepatocarcinoma." Thesis, Bordeaux 2, 2011. http://www.theses.fr/2011BOR21868/document.

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La polymérase NS5B du virus de l’hépatite C (VHC) porte une activité ARN polymérase ARN-dépendante essentielle pour la réplication de l'ARN génomique viral. Cette réplication implique la synthèse d'un intermédiaire de réplication de polarité négative. In vitro et probablement in vivo, la NS5B initie la synthèse d'ARN par un mécanisme de novo qui nécessite des interactions spécifiques entre la polymérase virale et des éléments des ARN viraux. Dans une première partie nous avons étudié le rôle du GTP et d’un domaine C-terminal nommé linker de la polymérase. Nos résultats démontrent que des concentrations élevées de GTP sont nécessaires pour la transition de l'initiation à l'élongation de la synthèse de l'ARN. Des mutations dans le linker à la position 556 ne modifient pas la concentration de GTP nécessaire pour la transition. Toutefois, l'initiation de la synthèse d'ARN est augmentée par la mutation S556K. Une analyse structurale menée en parallèle suggère une implication directe du linker dans l'initiation de novo de la synthèse de l'ARN. Dans les deuxièmes et troisièmes parties, nous avons étudié le rôle de motifs ARN dans la traduction et la synthèse de l’'ARN du VHC. Nous avons démontré que la tige boucle SL-E1 formée par la région entre les nt 177 et 222 de l'extrémité 3' de l’ARN (-) est importante pour la synthèse d'ARN in vitro par la NS5B recombinante et dans les cellules Huh7 exprimant le complexe de réplication (RC) du VHC. SL-E1 est impliquée dans l’initiation de la synthèse d’ARN, au moins in vitro. Nous avons également étudié le rôle des tiges boucles SLV et SLVI du gène core. Nos données n'ont pas montré de rôle évident de ces séquences ou de leur complément dans la synthèse de l'ARN in vitro par la NS5B recombinante et en culture cellulaire par le RC du VHC. Nous avons confirmé leur effet négatif sur la traduction IRES dépendante par interaction ARN-ARN longue distance entre SL-VI et le 5'UTR et démontré que le miR122 ne peut pas empêcher cet interaction. Par contre, la présence de SL-VI prévient l’inhibition de la traduction induite par l’interaction entre le domaine III de l’IRES et la tige boucle 5BSL3.2 en 3’ du génome. Ces résultats démontrent la complexité des interactions ARN/ARN et ARN/protéines dans la régulation de la réplication virale
The hepatitis C virus (HCV) NS5B protein displays a RNA-dependent RNA polymerase activity essential for replication of the viral RNA genome. This replication involves the synthesis of a replication intermediate of negative polarity. In vitro and likely in vivo, the NS5B initiates RNA synthesis by a de novo mechanism which requires specific interactions between the polymerase and viral RNA elements. In the first part of results, we described a combined structural and functional analysis of HCV-NS5B to study the role of a C-terminal segment (termed linker) and of GTP in RNA synthesis. Our results demonstrated that high GTP concentrations are necessary for the transition from the initiation to the elongation of RNA synthesis, and that linker mutations at position S556 did not modify the GTP requirement of NS5B for this transition. However, the initiation of RNA synthesis was greatly enhanced by a S556K mutation. These results together with a structural analysis point to the direct involvement of the linker in the de novo initiation of RNA synthesis. In the second and third parts of results, we studied the role of RNA elements in RNA synthesis. We demonstrated that the SL-E1 stem–loop formed by nucleotides 177–222 from the 3’-end of the HCV (-) RNA is important for RNA synthesis both in vitro by the recombinant NS5B and in Huh7 cells by HCV replication complex (RC). We also showed that SL-E1 is involved in initiation of RNA synthesis, at least in vitro. Then we studied the role of other viral RNA elements in core coding sequences (SLV and SLVI stem loops) and the involvement of the microRNA miR122 in RNA translation and RNA synthesis. For SLV and SLVI, our data did not show any clear role of these core-coding sequences or of their complement in the (-) RNA in RNA synthesis both in vitro by the recombinant NS5B and in cell culture by HCV-RC. We confirmed their negative effect on HCV-IRES translation through long range RNA-RNA interaction between SL-VI sequences and the 5’UTR and demonstrated that miR122 cannot disrupted this interaction and switches the region to an open conformation. Conversely, our data indicated that the SL-VI domain can counteract the negative effect of the interaction between the domain III of IRES and the 5BSL3.2 stem loop localized at the 3’end of the genome. These results point to the complexity of RNA/RNA and RNA/proteins interactions in the HCV replication cycle
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Books on the topic "Synthèss in vitro"

1

Conn, Graeme L., ed. Recombinant and In Vitro RNA Synthesis. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-62703-113-4.

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International Symposium on In Vitro Immunization in Hybridoma Technology (1987 Tylösand, Sweden). In vitro immunization in hybridoma technology: Proceedings of the International Symposium on In Vitro Immunization in Hybridoma Technology, Tylösand, Sweden, September 7-8, 1987. Amsterdam: Elsevier, 1988.

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Kawasaki, Arthur Jun. Investigations of in vitro and in vivo aspects of hepatic very low density lipoprotein synthesis and secretion in rat. Ottawa: National Library of Canada, 1995.

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Duncan, Sharon A. The isolation and characterization of two biliary cyclosporine A metabolites and an in vitro study of the inhibition of DNA synthesis of renal cells by cyclosporine A metabolites. Ottawa: National Library of Canada, 1990.

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Naumenko, V., B. Sorochynskyi, and Ya Blume. Synthesis of secondary metabolites in vitro. PH "Akademperiodyka", 2015. http://dx.doi.org/10.15407/akademperiodyka.281.056.

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Recombinant And In Vitro Rna Synthesis Methods And Protocols. Humana Press, 2012.

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International Symposium on In Vitro Immunization in Hybridoma Technology (1987 Tylösand, Sweden). In vitro immunization in hybridoma technology. Elsevier, 1988.

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F, Simmon Vincent, and Health Effects Research Laboratory (Research Triangle Park, N.C.), eds. In vitro microbiological mutagenecity and unscheduled DNA synthesis studies of fifteen pesticides. Research Triangle Park, NC: U.S. Environmental Protection Agency, Health Effects Research Laboratory, 1985.

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In Vitro Transcription and Translation Protocols. 2nd ed. Humana Press, 2007.

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J, Tymms Martin, ed. In vitro transcription and translation protocols. Totowa, N.J: Humana Press, 1995.

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Book chapters on the topic "Synthèss in vitro"

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Matthews, Jayne A., and Raymond A. McKee. "In Vitro Protein Synthesis." In Springer Protocols Handbooks, 131–47. Totowa, NJ: Humana Press, 1986. http://dx.doi.org/10.1007/978-1-60327-405-0_14.

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Bégin, R., and B. T. Mossman. "Summary and Synthesis of Part IV. Synthesis and Release of Mediators (A)." In In Vitro Effects of Mineral Dusts, 383. Berlin, Heidelberg: Springer Berlin Heidelberg, 1985. http://dx.doi.org/10.1007/978-3-642-70630-1_48.

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Hou, Ya-Ming. "High-Purity Enzymatic Synthesis of Site-Specifically Modified tRNA." In Recombinant and In Vitro RNA Synthesis, 195–212. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-62703-113-4_15.

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Booy, Evan P., Hui Meng, and Sean A. McKenna. "Native RNA Purification by Gel Filtration Chromatography." In Recombinant and In Vitro RNA Synthesis, 69–81. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-62703-113-4_6.

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Avis, Johanna M., Graeme L. Conn, and Scott C. Walker. "Cis-Acting Ribozymes for the Production of RNA In Vitro Transcripts with Defined 5′ and 3′ Ends." In Recombinant and In Vitro RNA Synthesis, 83–98. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-62703-113-4_7.

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Szafraniec, Milena, Leszek Blaszczyk, Jan Wrzesinski, and Jerzy Ciesiolka. "Trans-Acting Antigenomic HDV Ribozyme for Production of In Vitro Transcripts with Homogenous 3′ Ends." In Recombinant and In Vitro RNA Synthesis, 99–111. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-62703-113-4_8.

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Cheong, Hae-Kap, Eunha Hwang, and Chaejoon Cheong. "Rapid Preparation of RNA Samples Using DNA-Affinity Chromatography and DNAzyme Methods." In Recombinant and In Vitro RNA Synthesis, 113–21. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-62703-113-4_9.

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Bowman, Jessica C., Bahareh Azizi, Timothy K. Lenz, Poorna Roy, and Loren Dean Williams. "Preparation of Long Templates for RNA In Vitro Transcription by Recursive PCR." In Recombinant and In Vitro RNA Synthesis, 19–41. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-62703-113-4_3.

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Linpinsel, Jo L., and Graeme L. Conn. "General Protocols for Preparation of Plasmid DNA Template, RNA In Vitro Transcription, and RNA Purification by Denaturing PAGE." In Recombinant and In Vitro RNA Synthesis, 43–58. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-62703-113-4_4.

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Lu, Cheng, and Pingwei Li. "Preparation of Short RNA by In Vitro Transcription." In Recombinant and In Vitro RNA Synthesis, 59–68. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-62703-113-4_5.

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Conference papers on the topic "Synthèss in vitro"

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Kuruma, Yutetsu, Hideaki Matsubayashi, and Takuya Ueda. "In Vitro Reconstruction of Functional Membrane." In Artificial Life 14: International Conference on the Synthesis and Simulation of Living Systems. The MIT Press, 2014. http://dx.doi.org/10.7551/978-0-262-32621-6-ch156.

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Allochio Filho, J. F., L. L. Roldi, R. Fiorot, S. J. Greco, V. Lacerda Jr., R. B. dos Santos, E. V. R. de Castro, et al. "Synthesis and in vitro antifungal activity of new Mannich bases derived from 2-hydroxy-1,4-naphthoquinone (Lawsone)." In 15th Brazilian Meeting on Organic Synthesis. São Paulo: Editora Edgard Blücher, 2013. http://dx.doi.org/10.5151/chempro-15bmos-bmos2013_2013916122532.

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Akama, Satoru, Masayuki Yamamura, and Takanori Kigawa. "Multi-Objective Robust Optimization for In Vitro RNA Synthesis." In Computational Intelligence and Bioinformatics / Modelling, Simulation, and Identification. Calgary,AB,Canada: ACTAPRESS, 2011. http://dx.doi.org/10.2316/p.2011.753-017.

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Akama, Satoru, Masayuki Yamamura, and Takanori Kigawa. "Multi-Objective Robust Optimization for In Vitro RNA Synthesis." In Computational Intelligence and Bioinformatics / Modelling, Simulation, and Identification. Calgary,AB,Canada: ACTAPRESS, 2012. http://dx.doi.org/10.2316/p.2012.753-017.

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Kuang, Baoping, Hui Zhang, and Zhihang Shang. "Synthesis of Histone Deacetylases Inhibitor and Activity in Vitro." In 5th International Conference on Information Engineering for Mechanics and Materials. Paris, France: Atlantis Press, 2015. http://dx.doi.org/10.2991/icimm-15.2015.103.

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Bánóczi, Zoltán, Márton Flórián, Erika Orbán, Ildikó Szabó, and Ferenc Hudecz. "Methotrexate Containing Oligopeptide Conjugates: Synthesis and in vitro Cytostatic Effect." In The Twenty-Third American and the Sixth International Peptide Symposium. Prompt Scientific Publishing, 2013. http://dx.doi.org/10.17952/23aps.2013.094.

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Caschera, Filippo, and Vincent Noireaux. "A Novel in Vitro Metabolic Scheme for the Construction of a Minimal Biological Cell." In Artificial Life 14: International Conference on the Synthesis and Simulation of Living Systems. The MIT Press, 2014. http://dx.doi.org/10.7551/978-0-262-32621-6-ch089.

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Tremoli, E., D. Caruso, P. Maderna, G. Galli, and R. Paoletti. "DIFFERENTIAL EFFECTS OF ORAL ADMINISTRATIONS OF ACETYL SALICYLIC ACID, INDOMETHACIN AND SODIUM SALICYLATE TO HUMAN SUBJECTS ON THE FORMATION OF 12-HYDROXYEICOSATETRAENOIC BY PLATELETS." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643395.

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The effects of a single oral administration of acetylsalicylic acid (ASA, 500 mg), indomethacin (Indo, 50 mg) and sodium salicylate (SA, 400 mg) to healthy volunteers on platelet formation of 12-hydroxyeicosatetraenoic acid (12-HETE) were evaluated. Blood was collected before, 2, 6 and 24 hours after drug administrations. Platelet rich plasma (PRP) samples were stimulated with 20 μg/ml collagen and 12-HETE levels were determined by selective ion monitoring. The effects of ASA on the same parameter were also evaluated in vitro in PRP and in washed platelet (WP) samples in the absence and in the presence of platelet poor plasma (PPP, 25-100%) or bovine serum albumin (BSA, 10-40 mg/ml). In subjects who ingested ASA, the formation of 12-HETE in PRP stimulated with collagen was significantly inhibited 2 and 6 hours after the drug administration. At 24 h 12-HETE synthesis tended to return toward basal values. In contrast the administration of a single dose of Indo or of SA did not significantly affect 12-HETE synthesis by stimulated PRP. ASA (3 mM) added in vitro inhibited 12-HETE formation in PRP but not in WP. In addition ASA inhibited 12-HETE synthesis in washed platelets resuspended either in PPP (25-100 %) or in BSA (10-40 mg/ml). It is concluded ASA, but not Indo or SA, orally administered to normal subjects inhibits 12-HETE synthesis in collagen stimulated PRP. The results obtained in vitro suggest that albumin and/or some albumin component may be responsible for the inhibitory effect of ASA on platelet 12-HETE syntesis.
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Pethó, Lilla, József Murányi, Nassima Kram, Gyöngyi Bökönyi, Gabriella Csík, and Gábor Mezó. "Synthesis and In Vitro Biological Effect of Gnrh-Protoporphyrin IX Conjugates." In 35th European Peptide Symposium. Prompt Scientific Publishing, 2018. http://dx.doi.org/10.17952/35eps.2018.317.

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Didychuk, Candice L., Pinhas Ephrat, Michelle Belton, and Jeffrey J. L. Carson. "Synthesis and in vitro cytotoxicity of mPEG-SH modified gold nanorods." In Biomedical Optics (BiOS) 2008, edited by Alexander A. Oraevsky and Lihong V. Wang. SPIE, 2008. http://dx.doi.org/10.1117/12.763765.

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Reports on the topic "Synthèss in vitro"

1

Freeman, J. In vitro synthesis and purification of PhIP-deoxyguanosine and PhIP-DNA oligomer covalent complexes. Office of Scientific and Technical Information (OSTI), December 1994. http://dx.doi.org/10.2172/98639.

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Hemscheidt, Thomas. Semi-Synthesis and In-Vitro Anticancer Evaluation of Derivatives of a New Microtubule Poison with a Taxol-Like Mechanism. Fort Belvoir, VA: Defense Technical Information Center, September 2002. http://dx.doi.org/10.21236/ada411590.

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Hemscheidt, Thomas K. Semi-Synthesis and In-vitro Anticancer Evaluation of Derivatives of a New Microtubule Poison with a Taxol-Like Mechanism. Fort Belvoir, VA: Defense Technical Information Center, September 2004. http://dx.doi.org/10.21236/ada469112.

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Hemscheidt, Thomas U. Semi-Synthesis and In-Vitro Anticancer Evaluation of Derivatives of a New Microtubule Poison with a Taxol-Like Mechanism. Fort Belvoir, VA: Defense Technical Information Center, September 2003. http://dx.doi.org/10.21236/ada419388.

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