Дисертації з теми "Luciferasi"
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Conti, Elena Eliana. "Crystal structure of firefly luciferase." Thesis, Imperial College London, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.244284.
Повний текст джерелаWalpole, C. S. J. "Active site probes for bacterial luciferase." Thesis, University of Sussex, 1985. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.356510.
Повний текст джерелаLin, Leo Yen-Cheng. "Flavin binding site in Vibrio harveyi Luciferase." Thesis, McGill University, 2004. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=85083.
Повний текст джерелаChan, Wai Shing. "Applications of the bacterial luciferin-luciferase system." HKBU Institutional Repository, 2012. https://repository.hkbu.edu.hk/etd_ra/1454.
Повний текст джерелаGupta, Rajat. "Firefly luciferase mutants as sensors of proteome stress." Diss., lmu, 2012. http://nbn-resolving.de/urn:nbn:de:bvb:19-150266.
Повний текст джерелаLasko, Daniel R. "On-line fermentation monitoring via recombinant firefly luciferase." Thesis, Massachusetts Institute of Technology, 1995. http://hdl.handle.net/1721.1/11125.
Повний текст джерелаAndrews, Thomas. "A novel dual-luciferase monitoring apparatus a thesis /." San Antonio : UTHSC, 2008. http://learningobjects.library.uthscsa.edu/cdm4/item_viewer.php?CISOROOT=/theses&CISOPTR=36&CISOBOX=1&REC=20.
Повний текст джерелаOliveira, Anderson Garbuglio de. "Estudo mecanístico da bioluminescência de fungos." Universidade de São Paulo, 2010. http://www.teses.usp.br/teses/disponiveis/46/46135/tde-08112010-093327/.
Повний текст джерелаThis thesis describes how in vitro light emission can be enzymatically obtained from the hot and cold extracts assay using different species of fungi, which also indicates a common mechanism of light emission for all these organisms. Kinetic data suggest a consecutive two-step mechanism and corroborate the 1960\'s enzymatic proposal of Airth and Foerster. Finally, using hot and cold extracts assay we were also able to purify and to determine the molecular weight of the fungal luciferin (298.1837 m/z). The isolated substance emits light enzymatically in vitro, whose light emission spectrum matches with the fungal bioluminescence one thus confirming that the substance is the fungal luciferin
Buriánková, Karolína. "Résistance ribosomique aux macrolides et leur effet sur la fidélité de traduction." Paris 11, 2003. http://www.theses.fr/2003PA112220.
Повний текст джерелаMacrolide antibiotics constitute a homogenous group of antibacterial agents produced by Streptomyces or related Actinobacteria. They inhibit protein synthesis in bacteria by binding to the 50S ribosomal subunit, preventing its assembly or inhibiting its function. In the first part of this thesis the effect of macrolides on translation accuracy was studied. We have used the reporter system based on Vibrio harveyi luciferase with a stop codon inserted in the proximal part of the luxB gene for the in vivo measurement of the nonsense codon readthrough. Erythromycin stimulated the leadthrough of the UAG stop codon and thus the decrease of the translation accuracy. This is in agreement with the hypothesis that macrolides influence the early stages of elongation process. The misreading effect of macrolides was confirmed by the study of global error frequencies using the 2-D gel electrophoresis of proteins. The second part deals with the intrinsic macrolide resistance of the Mycobacterium tuberculosis complex (MTC), generally attributed to the low permeability of the mycobacterial cell wall. However we have shown that a gene, whose product confers macrolide resistance by ribosome modification, was present in all members of the MTC. It was named ermMT (erm. 37). Part of the ermMT is deleted in some vaccinal strains of Mycobacterium bovis BCG, such as the Pasteur strain. The Pasteur strain was susceptible to macrolides, whereas MTC species were resistant to them. The expression of ermMT in the macrolide-sensitive Mycobacterium strains conferred macrolide resistance. Comparison of the resistance patterns and ribosomal affinity for erythromycin of Mycobacterium host strains expressing ermMT or other erm genes indicates that ermMT confers a type I resistance to macrolides, lincosamides and streptogramins, coiresponding to the mono-methylation of A2058 in 23S rRNA. Our results indicate that ermMT plays a major role in the intrinsic macrolide resistance of members of the MTC
Eriksson, Jonas. "Advancements in Firefly Luciferase-Based Assays and Pyrosequencing Technology." Doctoral thesis, KTH, Biotechnology, 2004. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-3708.
Повний текст джерелаPyrosequencing is a new DNA sequencing method relying on thesequencing-by-synthesis principle and bioluminometric detectionof nucleotide incorporation events. The objective of thisthesis was improvement of the Pyrosequencing method byincreasing the thermal stability of firefly luciferase, and byintroducing an alternative DNA polymerase and a new nucleotideanalog. Furthermore, the development of a new bioluminescentassay is described for the detection of inorganicpyrophosphatase activity.
The wild-type North American firefly(Photinus pyralis)luciferase is a heat-sensitiveenzyme, the catalytic activity of which is rapidly lost attemperatures over 30°C. Two strategies for increasing thethermostability of the enzyme are presented and discussed. Inthe first strategy, the solution thermodynamics of the systemis affected by osmolytes in such a way that heat-mediatedinactivation of the enzyme is prevented. In the secondstrategy, the enzyme is thermostabilized by mutagenesis. Bothstabilizing strategies can be utilized to allow bioluminometricassays to be performed at higher temperatures. For instance,both DNA polymerase and ATP sulfurylase activity could beanalyzed at 37°C.
The osmolyte strategy was successfully employed forincreasing the reaction temperature for the Pyrosequencingmethod. By increasing the reaction temperature to 37°Cunspecific signals from primer-dimers and 3-end loopswere reduced. Furthermore, sequencing of a challenging templateat 37°C, which previously yielded poor, non-interpretablesequence signals at lower temperatures was now possible.
Introduction of a new adenosine nucleotide analog,7-deaza-2-deoxyadenosine-5-triphosphate (c7dATP) reduced the inhibitory effect on apyraseobserved with the currently used analog,2-deoxyadenosine-5-O-(1-thiotriphosphate)(dATPαS).
Sequencing of homopolymeric T-regions has previously beendifficult with the exonuclease-deficient form of the DNApolymerase I large (Klenow) fragment. By using the DNApolymerase from bacteriophage T7, known as Sequenase, templateswith homopolymeric T-regions were successfully sequenced.Furthermore, it was found that the strand displacement activityfor both polymerases was strongly assisted if the displacedstrand had a 5-overhang. In contrast, the stranddisplacement activity for both polymerases was inhibitedwithout an overhang, resulting in reduced sequencingperformance in double stranded regions.
A firefly bioluminescent assay for the real-time detectionof inorganic pyrophosphatase in the hydrolytic direction wasalso developed. The assay is versatile and has a linearresponse in the range between 8 and 500 mU.
Key words:bioluminescence, osmolytes, glycine betaine,thermostability, firefly luciferase, inorganic pyrophosphatase,inorganic pyrophosphate, Pyrosequencing technology, secondaryDNA-structures, Sequenase, Klenow-polymerase, reaction rates,temperature, c7dATP, dATPαS.
Farkasova, Katarina. "Bioluminescence imaging of luciferase transgenes in tumor metastases models." Diss., lmu, 2011. http://nbn-resolving.de/urn:nbn:de:bvb:19-139409.
Повний текст джерелаZeng, Jiang. "Chemiluminescence and bioluminescence related to flavins and bacterial luciferase." Thesis, University of Huddersfield, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.387194.
Повний текст джерелаJathoul, Amit Paul. "Activity of firefly luciferase with 6'-amino-D-luciferin." Thesis, University of Cambridge, 2008. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.612384.
Повний текст джерелаOliveira, Jordana Cristina. "Desenvolvimento de estratégias alternativas para teste de fármacos: obtenção e caracterização de linhagens mutantes estáveis de Leishmania expressando luciferase." Universidade de São Paulo, 2014. http://www.teses.usp.br/teses/disponiveis/42/42135/tde-15122014-154829/.
Повний текст джерелаLeishmaniasis is caused by protozoan parasites in Brazil, the main causative species of cutaneous leishmaniasis are Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonenses. The treatment of leishmaniasis presents several difficulties, and the discovery of new active drugs for the treatment of leishmaniasis is therefore fundamental. The enzyme luciferase is a reporter widely used in screening tests for new drugs. This enzyme catalyzes the oxidation of luciferase in the presence of ATP emitting light that can be detected in cultured cells in vitro as well as in intact animals, using the technique of bioimaging. In this work, we sought to produce strains of L. (V.) braziliensis and L. (L) amazonenses expressing luciferase and characterize the behavior of these mutant strains in drug susceptibility tests and in in vitro and in vivo infections. Production of light was detected in mutants of both species, in all life cycle stages. Mutant strains were compared to their corresponding parental lines as to their growth pattern, infectivity and survival profile in macrophages and sensitivity to amphotericin B and tamoxifen. No significant differences were observed for these parameters. BALB/c mice infected with the luciferase expressing line of L. (L.) amazonenses developed lesions comparable to those in animals infected with the wild-type strain. The parasite load in these animals was quantified through bioimaging. The results obtained of this study indicate that the mutant parasites expressing luciferase can be used for drug susceptibility testing in vitro and in vivo, representing a methodological advance in this area of research.
Curry, Stephen. "The interactions of general anaesthetics with a bacterial luciferase enzyme." Thesis, Imperial College London, 1989. http://hdl.handle.net/10044/1/47396.
Повний текст джерелаStowe, Cassandra. "Development of firefly luciferase bioluminescence for in vivo optical imaging." Thesis, University College London (University of London), 2018. http://discovery.ucl.ac.uk/10041771/.
Повний текст джерелаGilfoyle, David J. "Properties and applications of the beetle Luciferases." Thesis, University of Sussex, 1993. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.357617.
Повний текст джерелаFalklöf, Olle. "Photochemical properties of phytochrome and firefly luciferase chromophores: A theoretical study." Licentiate thesis, Linköpings universitet, Beräkningsfysik, 2014. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-103338.
Повний текст джерелаMoss, Guy William John. "The interactions of general anaesthetics and high pressure with firefly luciferase." Thesis, Imperial College London, 1989. http://hdl.handle.net/10044/1/47575.
Повний текст джерелаLaw, Gim Hoong Erica. "Mutational analysis of solvent-exposed amino acids in Photinus pyralis luciferase." Thesis, University of Cambridge, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.615816.
Повний текст джерелаHalliwell, Lisa Marie. "Protein engineering utilising single amino acid deletions within Photinus pyralis firefly luciferase." Thesis, Cardiff University, 2015. http://orca.cf.ac.uk/89475/.
Повний текст джерелаLake, Madryn. "Non-invasive imaging of estrogen receptor-coregulator interaction by luciferase fragment complementation." Thesis, Imperial College London, 2012. http://hdl.handle.net/10044/1/9256.
Повний текст джерелаMok, Pui-Wing. "Design and Applications of Split-Luciferase Systems in Vitro and in Cellulo." Diss., The University of Arizona, 2014. http://hdl.handle.net/10150/333354.
Повний текст джерелаScott, Mark George Hunter. "Control of cyclic AMP-mediated and ß₂ adrenergic receptor gene expression in cultured human airway smooth muscle cells." Thesis, University of Nottingham, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.324123.
Повний текст джерелаKemp, Daniel M. "Reporter gene analysis of regulatory mechanisms in cAMP signalling." Thesis, University of Kent, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.310202.
Повний текст джерелаHildebrandt, Stefanie. "Etablierung des Luciferase-Reportergenassays zur Quantifizierung der Bioaktivität von Leptin in menschlichem Serum." Doctoral thesis, Universitätsbibliothek Leipzig, 2013. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-113565.
Повний текст джерелаGupta, Rajat [Verfasser], and Ulrich [Akademischer Betreuer] Hartl. "Firefly luciferase mutants as sensors of proteome stress / Rajat Gupta. Betreuer: Ulrich Hartl." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2012. http://d-nb.info/1028191898/34.
Повний текст джерелаMurray, Shane Louise. "Identification and characterisation of Arabidopsis systemic acquired resistance mutants isolated by luciferase imaging." Thesis, University of Edinburgh, 2000. http://hdl.handle.net/1842/11207.
Повний текст джерелаJester, Benjamin. "Development of a Three-Hybrid Split-Luciferase System for Interrogating Protein Kinase Inhibition." Diss., The University of Arizona, 2011. http://hdl.handle.net/10150/205450.
Повний текст джерелаWaldenmaier, Hans Eugene. "Bioluminescência fúngica: papel ecológico, purificação e clonagem de enzimas." Universidade de São Paulo, 2016. http://www.teses.usp.br/teses/disponiveis/46/46131/tde-14072017-145527/.
Повний текст джерелаThis PhD thesis describes the studies performed to elucidate the molecular biology of fungal bioluminescence and the ecological significance of the trait in the wild. The recent discovery that the fungal luciferin is 3-hydroxyhispidin has allowed for the characterization of phenylalanine secondary metabolism in the newly sequenced genomes and mycelium transcriptomes of luminescent Panellus stipticus and Neonothopanus gardneri, additionally the genomes and transcriptomes of a non-luminescent variety of P. stipticus and Lentinula edodes served as respective controls. In general the genes involved in phenylalanine secondary metabolism had greater or equal expression in luminescent samples than non luminescent. A cluster of genes related to the secondary metabolism of phenylalanine was found in both luminescent and non luminescent P. stipticus genomes. Transcript abundance of genes in this cluster was similar in both luminescent and non-luminescent Panellus stipticus, but the type I polyketide synthase in non luminescent Panellus stipticus was significantly down regulated. A similar gene cluster in the N. gardneri and L. edodes genomes was absent with corresponding homologues scattered at different genomic loci. Cell free fungal extracts can be combined in vitro with the addition of 3-hydroxyhispidin to produce abundant green light. Preparation of proteinaceous luciferase extracts was improved and partially purified luciferase samples were investigated by mass spectrometry. The presence of luciferase in the separation gel was also evidenced by using luciferin and luciferin-like molecules from plant extracts. The ecological niche surrounding bioluminescent mushrooms was investigated through two main means, glue traps with acrylic mushroom facsimiles that were internally illuminated with green LED lights and direct observation of bioluminescent mushrooms with infrared time lapse photography. Ecological studies were performed in the Atlantic rainforest (Mata Atlântica) and transitional Coconut Palm forest (Mata dos Cocais) biomes of Brazil. Cockroaches, spiders, earwigs, crickets, and luminescent click beetles were the most common animal interacting with mushrooms. All of these animals may be acting as fungal propagule dispersers and in some cases defense of the mushroom.
Maiwald, Gelja. "In vivo bioluminescence imaging for monitoring of siRNA mediated luciferase knockdown in tumor models." kostenfrei, 2009. http://d-nb.info/1001449320/34.
Повний текст джерелаMaiwald, Gelja. "In vivo bioluminescence imaging for monitoring of siRNA mediated luciferase knockdown in tumor models." Diss., lmu, 2010. http://nbn-resolving.de/urn:nbn:de:bvb:19-113028.
Повний текст джерелаWiencierz, Anne Maria. "Entwicklung eines Dual-Luciferase-Reportergen-Assays zum Nachweis der Induktion antioxidativer Enzyme durch Nahrungsbestandteile." Master's thesis, Universität Potsdam, 2008. http://opus.kobv.de/ubp/volltexte/2009/2790/.
Повний текст джерелаThe induction of antioxidative enzymes might be an opportunity to elevate the cellular antioxidative capacity and, thus, to prevent oxidative stress associated diseases (e. g. cardio-vascular disease, neurodegenerative disease, atherosclerosis). Based on this idea the dual luciferase reporter gene (DLR) assay was developed to demonstrate the induction of three antioxidative enzymes: catalase (CAT), cytosolic glutathione peroxidase (GPX1), and copper-zinc superoxide dismutase (SOD1). In the course of the development three mammalian cell lines (CaCo2, IEC-18, V79) were tested for their ability to serve as a model cell line. The line V79 was chosen due to the transfection efficiency. To give consideration to a high-throughput several parameters were studied (e. g. format of the cultural plates, amount of DNA, kinetics of the luciferases) and the DLR assay was successfully established in 96 well plates. Subsequently, L-carnitine, catechin, epigallocatechin gallate, genistein, hydrogen peroxide (H2O2), sodium ascorbate, paraquat, quercetin, 12-O-tetradecanoylphorbol-13-acetate (TPA) and trolox were tested in non-cytotoxic concentrations for the activation of the rat CAT, human GPX1 and human SOD1 promoter. The maximally tolerable concentrations were determined by resazurin test in advance. Three out of these ten compounds were identified as potent inducers of GPX1 and SOD1. Stimulation of reporter gene construct transient transfected V79 cells for 24 hours with 100 µM paraquat caused a duplication of the relative GPX1 promoter activity and a 1.6-/1.7-fold increase of the relative SOD1 promoter activity. The incubation with 20 µM gen-istein or 10 µM quercetin resulted in duplication to triplication of both, the relative GPX1 and SOD1 promoter activity. In contrast, the rat CAT promoter was activated by 50 µM H2O2 (1.5-fold). Consequently, genistein, quercetin, and H2O2 are considered to be suitable reference substances for this DLR assay. To further characterize the inducing potential of the tested compounds all of them should be tested in different concentrations. Furthermore, for the routinely performed DLR assay it is recommended to use stably transfected cells to eliminate transfection caused variations.
Cowan, Heather Elizabeth. "Rapid, Quantitative Assessment of Antimycobacterial Water Disinfection based on the Firefly Luciferase Reporter Gene." Thesis, Virginia Tech, 1997. http://hdl.handle.net/10919/9748.
Повний текст джерелаMaster of Science
Farkašová, Katarina [Verfasser], and Eckhard [Akademischer Betreuer] Wolf. "Bioluminescence imaging of luciferase transgenes in tumor metastases models / Katarína Farkašová. Betreuer: Eckhard Wolf." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2011. http://d-nb.info/1019479302/34.
Повний текст джерелаErber, Astrid Maria [Verfasser], and Andreas [Akademischer Betreuer] Bechthold. "Untersuchungen zu Luciferase-ähnlichen Monooxygenasen, Flavinreduktasen und Ketoreduktasen aus dem Mensacarcin-Produzenten Streptomyces bottropensis." Freiburg : Universität, 2017. http://d-nb.info/1135572178/34.
Повний текст джерелаTemperley, Richard James. "Generation of a reporter for mitochondrial gene expression studies." Thesis, University of Newcastle Upon Tyne, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.369782.
Повний текст джерелаCampbell, Zachary Taylor. "STUDIES ON THE MECHANISM OF BACTERIAL BIOLUMINESCENCE IN VIVO AND IN VITRO." Diss., The University of Arizona, 2009. http://hdl.handle.net/10150/195376.
Повний текст джерелаCabral, Priscilla Carvalho. "Desenvolvimento de modelo experimental murino para o estudo da imunobiologia do melanoma." Universidade de São Paulo, 2016. http://www.teses.usp.br/teses/disponiveis/42/42133/tde-16112016-093857/.
Повний текст джерелаCancer is characterized as a multifactorial disease responsible for many deaths globally. Although nowadays we can find positive perspectives regarding cancers treatment, it is still very common to notice some alarming data. Therefore, our group developed some genetically modified tumoral lineages expressing ovalbumin (mOVA or cOVA) together with luciferase, in order to elucidate the relationship between tumor and the immune system. In our results, the presence of ovalbumin demonstrated: Changes in tumoral growth when animals were previously immunized with OVA and then challenged with our tumoral lineages; TCD8+ lymphocytes anti-OVA activation thus ovalbumin immunogenic potential when lineages were exposed to necrotic death followed by in vivo administration. In summary, our model showed that anti-tumoral vaccinations are indeed capable of promoting immune systems activation and consequently, improving the anti-tumor immunity.
Pereira, Tatiana Araujo. "Purificação e caracterização de enzimas envolvidas na bioluminescência de fungos." Universidade de São Paulo, 2017. http://www.teses.usp.br/teses/disponiveis/46/46136/tde-12042018-105954/.
Повний текст джерелаThis work describes studies performed to purify and characterize enzymes responsible for the fungal bioluminescence. Also, it shows important data that contributes to understand the mechanism for bioluminescence reaction in fungi. First, we tried to isolate two enzymes suspected of being involved on fungal bioluminescence. Optimum activity parameters (pH and temperature) and kinetic behavior were investigated. However, the discovery that fungal luciferin is the hispidin derivative 3-hydroxyhispidin demanded adaptations in the project. First of all, concentrates efforts to luciferase purification was priority, since hydroxylase is not part of the bioluminescent system of fungi. Studies on the luciferase interaction with different substrates showed some promiscuity for the enzyme. The results indicated higher intensity of light from luciferin-luciferase reaction in alkaline solutions (pH ~ 8) at ~ 20 °C. The reaction in medium with 18O2 revealed that insertion of oxygen into the luciferin structure produces an intermediate whose decarboxylation generates oxyluciferin. In parallel, the in vitro synthesis of hispidin using caffeic acid and malonyl-CoA with the mycelium extract resulted in the emission of light, confirming that luciferin is recycled in the process.
Gouilleux, Fabrice. "Facteurs impliques dans la regulation du promoteur du virus murin de la tumeur mammaire." Paris 11, 1991. http://www.theses.fr/1991PA115005.
Повний текст джерелаRoy, Karine. "Etude de la physiologie de Lactococcus lactis implanté dans le tractus digestif de souris par une approche protéomique." Paris 11, 2007. http://www.theses.fr/2007PA112050.
Повний текст джерелаKnowledge of the bacterial functions required for colonization of the digestive tract (DT) is essential for the understanding of the properties of the microbiote. In this thesis, we studied the adaptation of Lactococcus lactis to the DT of monoassociated mice. L. Lactis is one the main starter used in the dairy industry and recent developments have demonstrated its potential as a living vehicle for the targeting of antigens or therapeutics. To identify the functions expressed by in the DT we used monoassociated animal model (mice) combined with a proteomic approach. L. Lactis is established at a population level equivalent to that of commensal bacteria. Proteome analysis indicated that the fermentation pathways activated in the DT by L. Lactis is reminiscent to that observed during carbon starvation. We identified YwcC, a protein of unknown function as essential for DT colonization. We showed that YwcC possesses a phosphogluconolactonase activity and is thus implicated in the pentose phosphate pathway. In a second study, we added lactose to the diet of L. Lactis monoassociated mice. The addition of the sugar induced the synthesis of lactose catabolic enzymes and repressed the synthesis of proteins involved in alternative catabolic pathways. This result confirmed the hypothesis of a carbon starvation physiological state for L. Lactis established in the DT. We observed that the presence of lactose provided a strong competitive advantage to strains able to catabolize it. By the combination of gnotobiotic animals, comparative proteome analysis and mutants construction, this thesis work provides novel informations on the in vivo physiology of Lactococcus lactis
Doran, Diane Michelle. "HYPOXIC INDUCTION AND THE ROLE OF HIFS IN THE ACTIVATION OF LUCIFERASE CONSTITUTIVE REPORTERS IN PLACENTAL STEM CELLS." Wright State University / OhioLINK, 2007. http://rave.ohiolink.edu/etdc/view?acc_num=wright1190149635.
Повний текст джерелаDi, Bonito Rita. "Use of LuxA sequences for investigation on Luciferases kinetics and characterization of luminous bacteria." FIU Digital Commons, 2002. http://digitalcommons.fiu.edu/etd/2796.
Повний текст джерелаRienzo, Alessandro. "Estudio de la regulación dinámica de la expresión génica en respuesta a estrés osmótico en levadura." Doctoral thesis, Universitat Politècnica de València, 2016. http://hdl.handle.net/10251/62160.
Повний текст джерела[ES] Las células responden a los estímulos ambientales a través de una regulación precisa de la expresión génica. En este trabajo se investigó la modulación dosis dependiente de la expresión de genes activados en respuesta a estrés y por nutrientes. Se utilizó una versión desestabilizada de luciferasa de luciérnaga en células vivas de levadura como reportero para la detección de la expresión génica en tiempo real. Este sistema altamente sensible y no invasivo puede ser utilizado simultáneamente en diferentes condiciones experimentales a través de pequeñas alícuotas de cultivo. Esto permite la caracterización dosis-respuesta de la regulación de los promotores de levadura y puede ser utilizado para cuantificar parámetros importantes como el umbral, la sensibilidad, el tiempo de respuesta, la actividad máxima y el ratio de síntesis provocado por un determinado estímulo. Se aplicó el ensayo luciferasa al promotor GAL1 regulado por nutrientes y al promotor GRE2 activado en respuesta a estrés. Se observó que la expresión de la luciferasa activada por el promotor GAL1 responde de forma dinámica a las crecientes concentraciones de galactosa, con un incremento del ratio de síntesis determinado por el aumento de luz en la fase lineal inicial de la activación, en función de una gama de concentraciones de galactosa bien definidas. Este mecanismo de regulación depende de un aumento en la remodelación de las histonas y la consecuente asociación del complejo ARN pol II. La remodelación de la cromatina dosis dependiente parece ser la base de la expresión dinámica de GAL1, pues los mutantes relacionados con la dinámica de las histonas muestran perfiles dosis respuesta severamente afectados. En el caso del promotor GRE2, se demostró que una versión de una luciferasa desestabilizada es una herramienta excelente para describir de forma cuantitativa la activación transcipcional transitoria. La expresión de la luciferasa controlada por el promotor GRE2 responde de forma dinámica al aumento gradual de estímulo de estrés osmótico u oxidativo. La activación se observa principalmente en el incremento progresivo del tiempo en que el promotor permanece activo. Diferentemente de GAL1, el promotor GRE2 opera a través de un cambio apagado/encendido en respuesta a un aumento de estrés osmótico a través de ratios de síntesis prácticamente constantes y cuya regulación solamente depende de la remodelación de la cromatina y de la permanencia de la ARN pol II. Finalmente, se puede especular que el inductor Gal3 y la MAPK Hog1 son las moléculas determinantes para las diferentes estrategias de respuesta dinámica para los dos promotores. En este trabajo se identifican importantes diferencias en la señalización dinámica determinada por la dosis de estímulo en la expresión génica. En conjunto, el ensayo de luciferasa presentado en este trabajo puede ser una herramienta interesante para determinar y comparar de forma rápida y precisa los parámetros de la expresión génica. Adicionalmente se investigó la función del factor de transcripción Smp1 involucrado en la respuesta a osmoestrés en levadura. Un análisis de la asociación a la cromatina in vivo bajo estrés osmótico demostró que Smp1 se une preferentemente a regiones transcritas (ORFs) lo que refleja un comportamiento diferente comparando con otros activadores transcripcionales de la respuesta a estrés osmótico. Sin embargo, Smp1 parece ser importante para la expresión génica activada por estrés osmótico sólo en la presencia del gen natural inducido y no de fusiones artificiales del promotor. Esto evidencia el posible papel de Smp1 en la regulación de la expresión génica desde secuencias ORF y no en las regiones promotoras.
[CA] Les cèl·lules responen als estímuls ambientals a través d'una regulació precisa de l'expressió gènica. A aquest treball es va investigar la modulació dosi dependent de l'expressió de gens activats en resposta a estrès i per nutrients. Es va utilitzar una versió desestabilitzada de luciferasa de cuca de llum en cèl·lules vives de llevat com a reporter per a la detecció de l'expressió gènica a temps real. Aquest sistema altament sensible i no invasiu pot ser utilitzat simultàniament en diferents condicions experimentals a través de xicotetes alíquotes de cultiu. Això permet la caracterització dosi-resposta de la regulació dels promotors de llevat i pot ser utilitzat per a quantificar paràmetres importants com el llindar, la sensibilitat, el temps de resposta, l'activitat màxima i el rati de síntesi provocat per un determinat estímul. L'assaig luciferasa es va aplicar al promotor GAL1 regulat per nutrients i al promotor GRE2 activat en resposta a estrès. Es va observar que l'expressió de la luciferasa activada pel promotor GAL1 respon de forma dinàmica a les creixents concentracions de galactosa, amb un increment del rati de síntesi determinat per l'augment de llum en la fase lineal inicial de l'activació, en funció d'una gama de concentracions de galactosa ben definides. Aquest mecanisme de regulació depèn d'un augment en la remodelació de les histones i la conseqüent associació del complex ARN pol II. La remodelació de la cromatina dosi dependent sembla ser la base de l'expressió dinàmica de GAL1, ja què els mutants relacionats amb la dinàmica de les histones mostren perfils dosi-resposta severament afectats. En el cas del promotor GRE2, es va demostrar que una versió d'una luciferasa desestabilitzada és una eina excel·lent per a descriure de forma quantitativa l'activació transcripcional transitòria. L'expressió de la luciferasa controlada pel promotor GRE2 respon de forma dinàmica a l'augment gradual d'estímul d'estrès osmòtic o oxidatiu. L'activació s'observa principalment a l'increment progressiu del temps al qual el promotor roman actiu. De forma diferent de GAL1, el promotor GRE2 opera a través d'un canvi apagat/encès en resposta a un augment d'estrès osmòtic a través de ratis de síntesi pràcticament constants i als quals la seua regulació només depèn de la remodelació de la cromatina i de la permanència de l'ARN pol II. Finalment, es pot especular que l'inductor Gal3 i la MAPK Hog1 són les molècules determinants per a les diferents estratègies de resposta dinàmica per als dos promotors. A aquest treball s'identifiquen importants diferències a la senyalització dinàmica determinada per la dosi d'estímul a l'expressió gènica. En conjunt, l'assaig luciferasa presentat a aquest treball pot ser una eina interesant per a determinar i comparar de forma ràpida i precisa els paràmetres de l'expressió gènica. Addicionalment, es va investigar la funció del factor de transcripció Smp1 involucrat en la resposta a osmoestrès en llevat. Una anàlisi de l'associació a la cromatina in vivo sota l'estrès osmòtic va demostrar que Smp1 s'uneix preferentment a regions transcrites (ORFs), el qual reflecteix un comportament diferent comparant amb altres activadors transcripcionals de la resposta a estrès osmòtic. Tot i així, Smp1 sembla ser important per a l'expressió gènica activada per estrès osmòtic només en la presència del gen natural induït i no de fusions artificials del promotor. Això evidencia el possible paper de Smp1 en la regulació de l'expressió gènica des de seqüències ORF i no a les regions promotores.
Rienzo, A. (2016). Estudio de la regulación dinámica de la expresión génica en respuesta a estrés osmótico en levadura [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/62160
TESIS
Mofford, David M. "Pushing The Boundaries of Bioluminescence Using Synthetic Luciferins: A Dissertation." eScholarship@UMMS, 2015. http://escholarship.umassmed.edu/gsbs_diss/794.
Повний текст джерелаZallen, Jeremy Benjamin. "American Lucifers: Makers and Masters of the Means of Light, 1750-1900." Thesis, Harvard University, 2014. http://dissertations.umi.com/gsas.harvard:11460.
Повний текст джерелаHistory
JI, XIAOFEI, NAGAHIDE TAKAHASHI, ALEKSIC BRANKO, RYOKO ISHIHARA, TAKU NAGAI, AKIHIRO MOURI, SHINICHI SAITO, NOBUHISA MAENO, TOSHIYA INADA, and NORIO OZAKI. "AN ASSOCIATION BETWEEN SEROTONIN RECEPTOR 3B GENE (HTR3B) AND TREATMENT-RESISTANT SCHIZOPHRENIA (TRS) IN A JAPANESE POPULATION." Nagoya University School of Medicine, 2008. http://hdl.handle.net/2237/9647.
Повний текст джерелаKassar, Telissa da Cunha. "Construção e caracterização de vírus recombinante de febre amarela expressando o gene repórter da Gaussialuciferase." Centro de Pesquisas Aggeu Magalhães, 2013. https://www.arca.fiocruz.br/handle/icict/10367.
Повний текст джерелаFundação Oswaldo Cruz. Centro de Pesquisas Aggeu Magalhães. Recife, PE, Brasil
O vírus da febre amarela (YFV, Yellow Fever Virus), um arbovírus da família Flaviridae,é o agente causador da febre amarela (FA), uma doença aguda, febril, não contagiosa, hemorrágica e potencialmente fatal. O YFV é endêmico em regiões tropicais da América do Sul e África. Apesar de sua significância como um problema de saúde pública, muitos mecanismos moleculares da biologia do YFV, como replicação do genoma e patogênese viral ainda não foram bem compreendidos. Avanços em genética reversa viral tem permitido a elucidação de mecanismos da biologia e comportamento viral, bem como a construção de vetores vacinais e desenvolvimento de drogas antivirais. No presente trabalho, descrevemos a construção e caracterização de um vírus recombinante de FA expressando o gene repórter da Gaussialuciferase (GLuc). Utilizando o sistema de recombinação homóloga em levedura, o gene repórter da Proteína Fluorescente Amarela (YFP, Yellow Fluorescent Protein) do vírus recombinante YFV-YFP-DENV1linker, previamente construído em nosso laboratório, foi substituído pelo gene repórter GLuc. A construção foi confirmada por PCR. Os RNAs virais genômicos foram sintetizados in vitro, e posteriormente transfectados em células BHK-21.As células transfectadas foram avaliadas por imunofluorescência indireta e mensuração do gene repórter GLuc. Dois clones foram recuperados e caracterizados em cultivo celular. Nós acreditamos que este vírus repórter deverá ser útilna triagem e desenvolvimento de drogas antivirais específicas, estudos de replicação virale competência vetorial, além da possível utilização como vetor viral vacinal
Felixberger, Johannes [Verfasser], and Armin [Akademischer Betreuer] Buschauer. "Luciferase complementation for the determination of arrestin recruitment: Investigations at histamine and NPY receptors / Johannes Felixberger. Betreuer: Armin Buschauer." Regensburg : Universitätsbibliothek Regensburg, 2016. http://d-nb.info/1082128015/34.
Повний текст джерела