Tesis sobre el tema "Kinetics of mRNAs expression"
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BENDER, Cecilia. "Study of Human T-Lymphotropic Virus type 2 mRNA kinetics of expression and identification of a novel splicing site". Doctoral thesis, Università degli Studi di Verona, 2010. http://hdl.handle.net/11562/343864.
Texto completoThe analysis of HTLV expression during the infection process is essential to understand the influence of viral gene products on proliferation, cell cycle and signalling. Recent studies carried out on HTLV-1-infected cells have provided information on the relative abundance and timing of expression of different viral transcripts. In the case of HTLV-2, very little information has been so far obtained on the profile of viral gene expression in infected cells, and the kinetics of transcript expression have not yet been investigated. The aim of the study was to further investigate HTLV-2 transcripts expression, by developing new quantitative analyses to measure the levels of expression of different HTLV-2 mRNAs. In particular the kinetics of HTLV-2 transcripts at different stages of virus gene expression and their quantitation in infected cell lines and in PBMCs from infected subjects, have been evaluated. Similarly to other retroviruses, HTLV-2 expresses multiple gene products from the same coding region by choosing different strategies, including a complex pattern of alternative splicing. HTLV-2 expression produces three major classes of mRNAs: unspliced mRNA for Gag, Protease and Pol proteins; singly spliced mRNAs for Env and accessory proteins p28 and p22/p20; and doubly spliced mRNAs for the regulatory Tax, Rex and the accessory p10, p11 and 1-2-B proteins. More recently, the new APH-2 protein coded by the negative strand of HTLV-2, and possibly involved in the transcriptional silencing of the virus in infected cells was described. The expression profile and kinetics of the different transcripts were analysed by RT-PCR assays using a series of splice-junction-specific primers and probes. The time courses of each HTLV-2 transcript in three different cellular systems, namely stably infected cells lines, transiently transfected cells and ex-vivo IL-2 stimulated PBMCs from infected subjects were investigated. The results obtained led to the quantitation of all known HTLV-2 mRNAs. Preliminary data showed different levels of env transcript in HTLV-2 subtypes A and B. Evidence for a differential env expression in HTLV-2B stably infected cells, prompted further investigation on the complex pattern of splicing between exons 1 and 2 and allowed the identification of a novel 3’ acceptor site (SS) of splicing. This novel 3’SS is used to express alternative spliced isoforms within the pX terminal region of HTLV-2, including the singly spliced env and the three doubly spliced bicistronic tax/rex, p10/p11 and 1-2-B transcripts. Results demonstrated that the novel 3’SS was utilised in both HTLV-2 subtypes, and that the novel env isoform, named env 1-2b, was preferentially expressed in 2B subtypes, while 2A subtypes used more efficiently the canonical splice site. The kinetics of total mRNA HTLV-2 expression in these three cellular systems presented a general pattern characterised by an initial low transcript level and a subsequent increase to reach a peak of expression after 21-24 hours in culture. The full length gag/pol mRNA was the most abundant one during the time course, and behaved as a late gene that peaked after 21 to 48 hours. The canonical env transcript was expressed at very low rates in stably infected cells, and it was undetectable in PBMCs from infected subjects and in cells transfected with the HTLV-2 proviral clone. By contrast, env 1-2b isoform was efficiently expressed in stably infected cells as well as in ex-vivo infected PBMCs, behaving as a late gene showing a gradual and steady increase in copy number and reaching maximum expression at 21 to 48 hours. The regulatory tax/rex gene was expressed early and with a high/intermediate transcription rate in HTLV-2B subtypes in both stably infected cells and ex-vivo PBMCs. In this study the kinetics of expression of the yet unkwnon 1-2-B gene was investigated and found to be expressed at high levels at early time points, whereas the doubly spliced mRNA of accessory p10/p11 genes were poorly expressed or under detection limit. Among the singly spliced mRNAs coding for other accessory proteins, the p28, p22/p20-II isoform was found to be highly expressed in both HTLV-2 A and B subtypes as compared to its alternative p28, p22/p20-I form. The APH-2 negative transcript was efficiently expressed at high levels in both stably infected and transfected cells and behaved as a late gene. However, in ex-vivo PBMCs its expression level and kinetics pattern appeared to be variable and a clear pattern of expression was not assessed. In conclusion, this study demonstrated that HTLV-2 transcripts of both A and B subtypes are differentially expressed. A temporal pattern of mRNA production, with early, tax/rex and 1-2-B and late, gag/pol, env, p28,p22/p20-II genes was established. The tax/rex early expression indicates that this protein is necessary at the beginning of the viral cycle to transactivate and regulate viral and cellular genes. Most structural genes were expressed late when the transcription of early genes was already decreasing, thus showing that a temporal switch was occurring between early to late genes production. This study also provided new clues on the selective use of alternative 3’SS within the pX region of HTLV-2 for both subtypes A and B. Overall these findings indicate that the control of HTLV-2 viral gene expression is highly regulated both in its kinetics and expression. Moreover, it suggests that the use of multiple acceptor sites might play an important role on the preferential expression of specific proteins in the different phases of the viral cycle.
Taylor, David C. "SELEX targeting mRNAs : the hunt for novel riboregulators /". free to MU campus, to others for purchase, 2001. http://wwwlib.umi.com/cr/mo/fullcit?p3013032.
Texto completoMacdonald, Murdo. "Expression of mRNAs encoding FMRFamide-related peptides in adult and embryo Helix aspersa". Thesis, University of St Andrews, 1993. http://hdl.handle.net/10023/14454.
Texto completoDrummond, D. R. "The stability, movement and expression of natural and synthetic mRNAs injected into Xenopus oocytes". Thesis, University of Warwick, 1985. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.373064.
Texto completoChang, Chih Chien Anne. "Developmental expression of GABAA[subscript]/BZ receptor subunit mRNAs in olivocerebellar circuitry of the mouse /". The Ohio State University, 1994. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487848891512785.
Texto completoRende, Francesca. "Kinetics and regulation of HTLV-1 gene expression". Doctoral thesis, Università degli studi di Padova, 2011. http://hdl.handle.net/11577/3421976.
Texto completoRIASSUNTO Il virus T-linfotropico umano di tipo 1 (HTLV-1) è l’agente eziologico di due distinte patologie, la leucemia/linfoma a cellule T dell’adulto (ATLL, adult T-cell leukemia/lymphoma), un'aggressiva neoplasia a carico dei linfociti T CD4+ maturi, e della paraparesi spastica tropicale/mielopatia associata ad HTLV-1 (TSP/HAM, tropical spastic paraparesis/HTLV-1-associated myelopathy), una patologia degenerativa del sistema nervoso centrale. La strategia di espressione genica di HTLV-1, caratterizzata dalla produzione di trascritti a partire da promotori localizzati sia nel filamento positivo che in quello negativo del genoma virale, da splicing alternativo e da traduzione bicistronica, incrementa notevolmente la capacità codificante di HTLV-1, con la conseguente espressione di numerosi geni regolatori ed accessori (Tax, Rex, p12, p13, p21rex, p30tof e HBZ) in aggiunta alle proteine strutturali e agli enzimi associati al virione, comuni a tutti i retrovirus (Gag, Pro, Pol ed Env). Nonostante oltre 30 anni di studi, diversi aspetti chiave del ciclo vitale di HTLV-1 e della sua patogenicità rimangono tutt'oggi non noti. In particolare, non è ancora chiaro se l'espressione genica di HTLV-1 sia caratterizzata da stadi di latenza, se i diversi geni virali presentino cinetiche di espressione distinte e quali meccanismi molecolari possano controllare questi fenomeni. Gli studi descritti nella presente tesi sono stati mirati a comprendere questi aspetti della regolazione genica di HTLV-1. A questo scopo abbiamo sviluppato un protocollo di Real Time RT-PCR associato all'impiego di primer specifici per le diverse giunzioni di splicing al fine di quantificare i diversi trascritti codificati da HTLV-1 e di analizzarne le cinetiche di espressione sia in cellule mononucleate di sangue periferico isolate da individui infettati con HTLV-1, che in cellule trasfettate con cloni molecolari di HTLV-1. I risultati ottenuti indicano che l'espressione degli mRNA codificati da HTLV-1 segue una precisa cinetica dopo riattivazione dell'espressione virale: l'mRNA codificante le proteine regolatrici Tax e Rex agisce come trascritto precoce che precede l'espressione degli altri geni virali. Sebbene sia comunemente accettato che Rex eserciti la sua funzione a livello post-trascrizionale controllando l'esporto nucleare e la stabilità degli mRNA che codificano le proteine associate al virione, fino ad oggi non è mai stata investigata la Rex-dipendenza dei trascritti p12, p13, p21rex, p30tof e hbz. Al fine di testare se le cinetiche di espressione genica di HTLV-1 osservate potessero dipendere dalla funzione di Rex e al fine di determinare la Rex-dipendenza dei singoli mRNA virali, abbiamo generato un clone molecolare di HTLV-1 knock-out per Rex e analizzato la compartimentalizzazione nucleo-citoplasmatica dei trascritti virali. I risultati ottenuti hanno dimostrato la stretta Rex-dipendenza delle cinetiche di espressione a "due fasi" ed hanno rivelato una forte ritenzione nucleare degli mRNA codificanti HBZ, supportando la loro funzione come trascritti non codificanti. Inoltre, i risultati ottenuti hanno dimostrato che la responsività a Rex dei differenti mRNA virali potrebbe essere determinata dalla presenza di una sequenza regolatoria di 72 nucleotidi che agisce in cis, localizzata a monte dell'esone 3. Infine, analisi matematiche hanno sottolineato l'importanza di un ritardo temporale tra le funzioni di Tax e di Rex, supportata dall'evidenza sperimentale di un ritardo nell'accumulo e di un'emivita più prolungata di Rex rispetto a Tax. I dati ottenuti in questo studio forniscono l'evidenza di una regolazione temporale dell'espressione genica di HTLV-1, rivelano una differente compartimentalizzazione degli mRNA virali e offrono una possibile spiegazione di un paradosso ancora irrisolto della regolazione di HTLV-1, ovvero la differente Rex-dipendenza dei trascritti virali, nonostante la presenza della sequenza responsiva a Rex (RxRE, Rex-responsive element) nella regione 3' non tradotta di tutti i trascritti virali.
Crutchfield, Gerald L. "Kruppel-Like Transcription Factor 6 & 7 mRNAs (KLF6 & KLF7) Expression in the Developing Zebrafish". University of Akron / OhioLINK, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=akron1572200378181869.
Texto completoGuo, Fang. "Regulation of expression of SSTR1 and SSTR2 somatostatin receptor mRNAs in the arcuate nucleus by growth hormone". Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1995. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp04/MQ37318.pdf.
Texto completoOhmori, Sachiko, Kazumi Kanda, Hirohito Mitsuyama, Takashi Miyazaki, Xia Cao, Fukushi Kambe y Hisao Seo. "Tail-suspension Induces Altered Expression of GAPDH and ZAKI-4β mRNAs in Rat Hindlimbs : Implication for Disuse Muscle Atrophy". Research Institute of Environmental Medicine, Nagoya University, 2003. http://hdl.handle.net/2237/7561.
Texto completoParkin, Neil T. "Regulation of gene expression by the 5' untranslated region of eukaryotic mRNAS : c-myc and HIV-1 as examples". Thesis, McGill University, 1989. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=74327.
Texto completoDonovan, Benjamin Thomas. "Nucleosome Regulation of Transcription Factor Binding Kinetics: Implications for Gene Expression". The Ohio State University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=osu1574774626880568.
Texto completoPignata, Luiz Fernando Martins. "Pipeline para Análise In Sílico de Dados de Expressão de miRNAs e mRNAs em Células de Mamíferos". Universidade de São Paulo, 2012. http://www.teses.usp.br/teses/disponiveis/17/17135/tde-14062012-132754/.
Texto completoThe microRNAs are involved in the regulation of gene expression of the cell. The miRNA molecule binds to the messenger RNA and interrupts the gene expression by disrupting the translation. Through microarray data analysis, bioinformatics is a valuable aid for the identification of several genes that encode miRNAs in plants and animals, including mammals. It is also very useful for predicting structures. Data of miRNA and mRNA expression were obtained by the collaboration the Bioinformatics Laboratory and the Molecular Immunogenetics Laboratory of the Department of Genetics of the Faculty of Medicine of Ribeirão Preto - USP, coordinated by professors Silvana Giuliatti and Geraldo A. S. Passos, respectively. During the development and tests of the research, microarrays data (numerical values os the expression) were obtained from the comparison between the expression of miRNA and mRNA of the thymus of non obese diabetic mice with diabetes mellitus type 1, as well as from comparisons of their expression in other experiments. The present study is aimed at the development of a pipeline for in silico analysis of the data of miRNAs and mRNA gene expression obtained by microarray. Based on miRNAs and mRNA expression, it was possible to analyze several tools, develop and adjust scripts that allowed the sequential analysis of such data. The pipeline includes the quantification of gene expression data from microarray, the normalization of the data, the statistical analysis of differentially expressed sequences using Multi Experiment Viewer, the construction of networks of interaction of miRNA-mRNAs, and the search for targets of miRNAs based on such network using GenMir++. The pipeline was performed easily and allowed the correct analysis of the data, avoiding waste of time in bench analysis. From the results, new targets of miRNA were found using the pipeline and were verified further in bench analysis. The results were presented in the 55 th Brazilian Genetics Congress in the paper entitled \"MicroRNA-mRNA Network Controlling the Promiscuous Gene Expression in the Thymus of NOD (Non Obese Diabetic) Mice: Implications in the Emergence of Type 1 Diabetes Mellitus\".
Fleeger, Melissa. "Expression kinetics of the quinic acid (qa) gene cluster in Neurospora crassa". Youngstown State University / OhioLINK, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1299165877.
Texto completoOliveira, Gabriella Borba de. "Integrative analysis of microRNAs and mRNAs involved in regulation of intramuscular fat deposition in Nelore cattle". Universidade de São Paulo, 2017. http://www.teses.usp.br/teses/disponiveis/11/11139/tde-26072017-103507/.
Texto completoA quantidade de gordura intramuscular pode influenciar as características sensoriais e o valor nutricional da carne bovina, assim, a seleção de animais com conteúdo de gordura adequado para o consumidor torna-se importante. A gordura intramuscular é uma característica complexa, de difícil medição e há um conhecimento crescente sobre os genes e vias que controlam os processos biológicos envolvidos na deposição de gordura no músculo. MicroRNAs (miRNAs) são uma classe bem conservados de pequenos RNAs não-codificantes, que modulam a expressão gênica de uma gama de funções no desenvolvimento e fisiologia animal. Este estudo objetivou identificar miRNAs diferencialmente expressos (DE), genes reguladores candidatos e redes de co-expressão usando dados de expressão de mRNAs e miRNAs do músculo Longissimus dorsi de 30 novilhos Nelore com valores genéticos genômicos estimados (GEBV) extremos para conteúdo de gordura intramuscular (IMF). A análise de expressão diferencial entre os dados de miRNA de animais com valores extremos de GEBV para o IMF identificou seis miRNAs DE. A anotação funcional de genes alvos destes microRNAs indica que a via de sinalização de PPAR está envolvida com a deposição de IMF. Os genes reguladores candidatos, tais como SDHAF4, FBXO17, ALDOA e PKM foram identificados pelas abordagens de correlação parcial com teoria da informação (PCIT), fator de impacto fenotípico (PIF) e fator de impacto regulatório (RIF) a partir de dados integrados de expressão de mRNAs-miRNAs. Dois miRNAs, bta-miR-143 e bta-miR-146b, com alta expressão no grupo de baixo conteúdo de IMF, também foram correlacionados com genes reguladores candidatos, os quais foram funcionalmente enriquecidos para termos GO relacionados a oxidação de ácidos graxos. As redes de co-expressão identificaram vários módulos relacionados ao sistema imunológico, ao metabolismo das proteínas, ao metabolismo energético e ao catabolismo da glicose através da análise ponderada da rede de correlação (WGCNA), que mostrou possível interação e regulação entre mRNAs e miRNAs. Este estudo contribui com a compreensão dos possíveis mecanismos reguladores das redes de sinalização genética envolvidas no processo de deposição de gordura. O metabolismo da glicose e o processo de inflamação foram as principais vias encontrados na análise integrada de mRNA-miRNA e mostraram estar associadas ao conteúdo de gordura intramuscular em bovinos de corte.
Yamaguchi, Shun. "Regional Expression and Regulation of Alternative Forms of mRNAs Derived from Two Distinct Transcription Initiation Sites of the Rat mGluR5 Gene". Kyoto University, 1998. http://hdl.handle.net/2433/182240.
Texto completoYamada, Lixy. "Embryonic expression profiles and conserved localization mechanisms of pem-like mRNAs of two species of ascidian, Ciona intestinalis and Ciona savignyi". 京都大学 (Kyoto University), 2006. http://hdl.handle.net/2433/144235.
Texto completoJiang, Fei. "Expression of recombinant manganese peroxidase in Pichia pastoris Biosynthesis, stability and kinetics analysis /". Related electronic resource: Current Research at SU : database of SU dissertations, recent titles available full text, 2006. http://proquest.umi.com/login?COPT=REJTPTU0NWQmSU5UPTAmVkVSPTI=&clientId=3739.
Texto completo吳欲勳 y Yuk-fun Ng. "Regulation of mammary tumor cell cycle kinetics and gene expression bydietary fatty acids". Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 1998. http://hub.hku.hk/bib/B31219779.
Texto completoJennings, Natalie A. "Acetylcholinesterase in the sea urchin Lytechinus variegatus characterization and developmental expression in larvae /". Birmingham, AL : University of Alabama at Birmingham, 2007. https://www.mhsl.uab.edu/dt/2008r/jennings.pdf.
Texto completoTouleimat, Mohamed Nizar. "Méthodologie d'extraction et d'analyse de réseaux de régulation de gènes : analyse de la réponse transcriptionnelle à l'irradiation chez S. cerevisiæ". Thesis, Evry-Val d'Essonne, 2008. http://www.theses.fr/2008EVRY0044/document.
Texto completoThe cellular response to the DNA damage provoked by irradiation (IR) is relatively well studied, however, many observations show the involvement of the expression of many genes. We propose to identify the different potential patterns of the transcriptional response to IR and to reconstruct a gene regulatory network involved in its control. The first point of this work lies in the exploitation of the gene expression dynamics in conditions of genetic perturbations. The second point lies in the integration of systemic biological informations. We define an approach composed of one step of automated logical deduction of regulations from a strategy of perturbations and two induction steps that allow the analysis of the gene expression dynamics and the extraction of potential regulation from additional data. This approach allowed to identify, for the yeast, a complex response to IR and allowed to propose a regulation model which some relations have been experimentally validated
Ng, Yuk-fun. "Regulation of mammary tumor cell cycle kinetics and gene expression by dietary fatty acids /". Hong Kong : University of Hong Kong, 1998. http://sunzi.lib.hku.hk/hkuto/record.jsp?B19471026.
Texto completoLawson, Vannice Rose Marie. "The molecular cloning and developmental expression of two novel mRNAs encoding putative 14 kDa ODF and FS proteins of the rat sperm tail". Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1998. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp04/mq28218.pdf.
Texto completoCalder, Michele D. "Ovarian cysts in dairy cattle : importance of serum LH concentrations in maintenance of cysts and expression of mRNAs for steroidogenic enzymes and gonadotropin receptors /". free to MU campus, to others for purchase, 1998. http://wwwlib.umi.com/cr/mo/fullcit?p9924869.
Texto completoAken, Alexander Frans Johan van. "Effects of the expression of alternative oxidase on oxidising pathway kinetics in Schizosaccharomyces pombe mitochondria". Thesis, University of Sussex, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.439148.
Texto completoDelcau, Michael Asher. "Differentiation of Pseudomonas sp. strain ADP biofilms and planktonic cells using methods in gene expression analysis". Diss., University of Iowa, 2018. https://ir.uiowa.edu/etd/6093.
Texto completoSATO, Jun, Kazue MIZUMURA, Ken TAKAHASHI, Kimiaki KATANOSAKA y Yasuko KOZAKI. "Increased C-fiber Activities to Cold in Adjuvant-monoarthritic Rats was not Accompanied by Increased Expression of TREK1 and CMR1 mRNAs(RIEM Conference II, 2002)". Research Institute of Environmental Medicine, Nagoya University, 2002. http://hdl.handle.net/2237/2809.
Texto completoGustafsson, Sofia. "Expression and Purification of Murine Tripeptidyl Peptidase II". Thesis, Uppsala universitet, Institutionen för medicinsk biokemi och mikrobiologi, 2012. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-177009.
Texto completoNaito, Hiroshi. "Altered expression of retinoic acid (RA) receptor mRNAs in the fetal mouse secondary palate by all-trans and 13-cis Ras : Implications for RA-induced teratogenesis". Kyoto University, 1999. http://hdl.handle.net/2433/181747.
Texto completoLuscieti, Sara. "The Iron Regulatory Protein/Iron Responsive Element (IRP/IRE) system: functional studies of new target mRNAs and pathological implications for novel IRE mutations". Doctoral thesis, Universitat de Barcelona, 2016. http://hdl.handle.net/10803/399593.
Texto completoEl hierro es un micronutriente esencial requerido por múltiples reacciones celulares y una regulación adecuada de su metabolismo es fundamental para la salud. La homeostasis celular del hierro está basada en la coordinación de la absorción, el almacenamiento y la movilización de este elemento. Estos procesos son controlados post-transcripcionalmente por el sistema IRP/IRE. Las proteínas reguladoras del hierro (IRP1 e IRP2) reconocen un motivo estructural presente en el ARNm de algunos genes denominado IRE (Iron Responsive Element). El IRE es un motivo conservado y situado en las regiones no traducidas (UTR) de los ARNm de proteínas implicadas en el metabolismo del hierro. Las interacciones IRP/IRE regulan la expresión de los ARNm que codifican proteínas para la adquisición, el almacenamiento, la utilización y la exportación de hierro en respuesta a los niveles celulares de hierro, porque la interacción de las IRPs con los motivos IRE se ve incrementada a niveles bajos de hierro y disminuida en condiciones altas de hierro. Según la localización de los IRE, las IRPs regulan de manera distinta la expresión de sus dianas: las IRPs inhiben la iniciación de la traducción cuando se unen a IREs situados en el 5' UTR, mientras que su asociación con los IREs del 3' UTR estabiliza y protege dicho ARNm de la degradación. La falta de coordinación de la expresión de genes que contienen IRE está asociada con condiciones patológicas ilustrando la importancia de los componentes del sistema regulador IRP/IRE. En las últimas décadas se han realizado progresos importantes en el campo del metabolismo del hierro. Sin embargo, la regulación post-transcripcional de la expresión génica por el sistema IRP/IRE se ha limitado a un pequeño subconjunto de genes. Un estudio “genome-wide” llevado a cabo en nuestro grupo para la caracterización global del sistema regulador IRP/IRE dio como resultado la identificación de 35 genes diana que se unen a IRP1 e IRP2. La presente tesis tiene como objetivo general validar y caracterizar funcionalmente una de esas dianas: Profilin2 (Pfn2). Pfn2 es una proteína que une actina y que está involucrada en la regulación de la dinámica del citoesqueleto. Hemos identificado un IRE localizado en el 3’ UTR del ARNm de Pfn2; el IRE es funcional en estudios in vitro para la unión a IRP1 e IRP2. El ARNm de Pfn2 viene preferentemente regulado y reducido en condiciones altas de hierro en líneas celulares y hemos demostrado in vivo que es regulado por estabilización mediada de las IRPs, ya que los niveles de ARNm de Pfn2 se encuentran reducidos en ratones a los que los genes de Irp1 y Irp2 han sido inactivados. Así mismo, la sobrexpresión de Pfn2 en líneas celulares asociada a una reducción en los niveles de hierro libre, así como la desregulación de la distribución del hierro encontrada en ratones “knockout” para el gen de Pfn2; convierten a Pfn2 como un actor nuevo en el metabolismo del hierro. Por otro lado hemos contribuido tanto a la identificación de un 3’ IRE funcional en el ARNm humano de BDH2, una proteína involucrada en el tráfico de hierro a través del sistema de lipocalinas-sideróforos, como a la identificación de dos nuevas mutaciones en el IRE de la Ferritina L (Heidelberg +52G>C y Badalona+36C>U) responsables del Síndrome Hereditario de Hiperferritinemia con Cataratas y también a la identificación de una nueva mutación en el IRE de ALAS2 que ha demostrado ser un modificador de la importancia clínica de los síntomas en una familia con Protoporfiria Eritropoyética.
Omlin, Teye D. "Effects of Hypoxia and Exercise on In Vivo Lactate Kinetics and Expression of Monocarboxylate Transporters in Rainbow Trout". Thèse, Université d'Ottawa / University of Ottawa, 2014. http://hdl.handle.net/10393/30652.
Texto completoRosario, Wenzel Ileana del. "Anatomical localization and kinetics of expression of HSV and MHC antigens during viral encephalitis in a mouse model /". The Ohio State University, 1993. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487842372895558.
Texto completoTourlet, Sébastien. "Proposition d’une stratégie d’analyse statistique des données de puces à ADN décrivant une cinétique d’expression génique". Thesis, Tours, 2009. http://www.theses.fr/2009TOUR3134.
Texto completoMicroarray results were blamed because of their lack of concordance. Moreover, the huge candidate gene lists from statistical filterings are not useful for biologists. FDA proved that the lack of reliability between microarray experiments came from the choice of gene filtering indicators. In this context, a filtering method was developed based on expression curve shape modelling with the use of Log 2 of fold-change between kinetic points. Actually, the co-regulated genes display similar expression shape but with heterogeneous expression level.Our method was developed and validated thanks to two independent microarray experiments (Affymetric®) from mouse embryonic ovaries. Therefore, a short and relevant list of genes was obtained. Thus, a study of results linked to ovarian differentiation permitted to identify nine new candidate genes that were in silico validated. These genes might be biologically tested (i.e. RT PCR) by the scientific community
Huang, Yehong. "The Kinetics of G2 and M Transitions Regulated by B Cyclins". Case Western Reserve University School of Graduate Studies / OhioLINK, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=case1386197228.
Texto completoTabata, Sumie. "Distribution and kinetics of SR-PSOX/CXCL16 and CXCR6 expression on human dendritic cell subsets and CD4[+] T cells". Kyoto University, 2006. http://hdl.handle.net/2433/143841.
Texto completoTakahashi, Paula. "Perfis de Expressão Gênica e Possíveis Interações entre microRNAs e mRNAs em Diabetes Mellitus Tipo 1 com Enfoque em Resposta ao Estresse Oxidativo e Reparo do DNA". Universidade de São Paulo, 2015. http://www.teses.usp.br/teses/disponiveis/17/17135/tde-01072015-092915/.
Texto completoType 1 Diabetes Mellitus (T1DM) results from an autoimmune attack against the pancreatic cells, ceasing insulin production, which causes hyperglycemia. Although associations between oxidative stress, which can cause DNA damage, and T1DM have been demonstrated, only a few studies have reported differential expression of genes associated with response to oxidative stress and DNA repair in T1DM patients. Moreover, microRNAs (post-transcriptional regulators of gene expression) are implicated in many biological processes and pathological conditions; however, only scarce information is available in the literature concerning the expression of microRNAs in T1DM. In order to better understand the regulatory pathways involved in biological processes that are relevant to T1DM, we aimed to investigate the microRNA and mRNA transcriptional expression profiles by microarray analysis (as well as expression of selected proteins) in peripheral blood mononuclear cells (PBMCs) from T1DM patients (n=19) compared with healthy non-diabetic individuals (n=11), emphasizing genes related to response to oxidative stress and DNA repair. Microarray expression results indicated 44 differentially expressed microRNAs (35 up- and nine down-regulated) in T1DM patients, with those microRNAs possessing a discriminatory power to clearly stratify the patients from the controls, including hsa-miR-101, hsa-miR148a, hsa-miR-27b, and hsa-miR-424, whose expression data were confirmed by qRT-PCR. Functional annotation analysis performed on the predicted targets of the differentially expressed microRNAs pointed 22 and 12 annotated KEGG pathways for the overexpressed and repressed microRNAs, respectively, many of them related to cancer. Regarding mRNA microarray results, we detected 277 differentially expressed genes in T1DM patients, with 52% of them being potential targets of the differentially expressed microRNAs in T1DM patients. Among these targets, we identified candidate genes for T1DM as well as genes involved in the biological processes response to oxidative stress and DNA repair, such as UCP3, PTGS2, ATF3, FOSB, DUSP1 and TNFAIP3, whose expression data were confirmed by qRT-PCR. Furthermore, out of the 49 and 55 significantly expressed/enriched gene sets in T1DM patients, respectively, five pathways related to apoptotic signaling, response to hydroperoxide, DNA repair via homologous recombination, and response to endoplasmic reticulum stress were of interest for the present work. Concerning protein expression results (western blotting), PTGS2 and ATF3 expression was not detected for either the patient or the control group, while significant difference in DUSP1 expression was not observed between the two groups, although the corresponding mRNAs of those genes were found induced. Regarding the luciferase assay, our results demonstrated that the interaction between hsa-miR-148a and DUSP1 occurs in the cellular milieu. Therefore, these findings together with those western blotting results suggest that hsa-miR-148a could play a role in DUSP1 translational repression. Altogether, our results indicate distinctive microRNA and mRNA expression profiles in PBMCs from T1DM patients relative to healthy non-diabetic individuals. Furthermore, we have provided novel data regarding microRNA-mRNA interactions in T1DM, in particular involving genes associated with response to oxidative stress and DNA repair, suggesting a perturbation in the microRNA-target network in T1DM patients.
Fernandez, Christine Cheryl. "Characterisation of cytochrome P450 azole drug-resistant sterol demethylase CYP51B1 and expression of CYP123 and CYP136 from Mycobacterium tuberculosis". Thesis, University of Manchester, 2011. http://www.manchester.ac.uk/escholar/uk-ac-man-scw:171502.
Texto completoBeloti, Lilian Luzia 1980. "Estudo funcional de uma epóxido hidrolase de Aspergillus brasiliensis CCT1435 = expressão, purificação e caracterização enzimática = Functional study of an epoxide hydrolase from Aspergillus brasiliensis CCT1435: expression, purification and enzymatic characterization". [s.n.], 2013. http://repositorio.unicamp.br/jspui/handle/REPOSIP/316511.
Texto completoTese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
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Doutorado
Genetica de Microorganismos
Doutora em Genética e Biologia Molecular
Lindås, Ann-Christin. "Tripeptidyl-Peptidase II : Structure, Function and Gene Regulation". Doctoral thesis, Uppsala University, Department of Biochemistry, 2006. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-7345.
Texto completoThe protein degradation process is of vital importance for the cell to maintain cellular functions. An important enzyme in this process is the multimeric tripeptidyl-peptidase II (TPP II). It removes tripeptides from a free N-terminus of the substrates. TPP II has broad substrate specificity and wide-spread distribution, suggesting that the TPP II gene is a house-keeping gene. However, the levels of both mRNA and TPP II protein varies during different conditions and the TPP II gene promoter was therefore identified and characterized. It is a 215 bp fragment just upstream of the coding sequence. This fragment lacks a TATA-box but contains an initiator, two inverted CCAAT-boxes and an E-box. The CCAAT-boxes and the E-box were found to bind the nuclear factor Y (NF-Y) and upstream stimulatory factor-1 (USF-1) respectively. The CCAAT-boxes appear to be most important for the transcriptional activation. Furthermore, several silencer element were identified further upstream of the 215 bp promoter and the octamer binding factor Oct-1 was found to bind one of these fragments. If Oct-1 is responsible for the inhibition of the transcription of the TPP II gene remains to be investigated. In addition, the substrate specificity was investigated. For this purpose an expression system using Pichia pastoris was developed. The purified recombinant TPP II was found to have the same enzymatic properties as the native enzyme. In order to identify the amino acids involved in the binding of the N-terminus of the substrate, wild-type murine TPP II and four mutants E305Q, E305K, E331Q and E331K were purified. Steady-state kinetic analysis clearly demonstrated that both Glu-305 and Glu-331 are important for this binding as the KMapp is more than 102 higher for the mutants than wild-type. Finally, the pH-dependence for cleavage of two chromogenic substrates was compared for TPP II from different species.
Marcaud, Hélène. "Etude sur les transitions coopératives et les cinétiques de transconformation associées dans les acides nucléiques". Paris 6, 1986. http://www.theses.fr/1986PA066514.
Texto completoYalala, Bongani Ndhlovu. "Ion exchange resins an functional fibres :a comparative study for the treatment of brine waste water". Thesis, University of the Western Cape, 2009. http://etd.uwc.ac.za/index.php?module=etd&action=viewtitle&id=gen8Srv25Nme4_8342_1298358875.
Texto completoTo improve the adsorption capacity of polyacrylonitrile (PAN) fibres, hydrophilic amidoxime fibres were prepared by subsequent conversion of the cyano groups to an amidoxime group by reacting with hydroxylamine at 80°
C at an optimum amidoximation time of 2 hrs. The amidoxime fibre was hydrolyzed/alkali treated in a solution of sodium hydroxide to enhance or improve the adsorption properties. This was followed by characterization of the amidoxime and hydrolyzed fibres using Scanning electron microscopy (SEM)
Fourier transform Infrared Spectroscopy (FTIR) and exchange capacity (cationic and anionic). SEM showed that the hydrolysis process made the surface of Amidoxime fibre rougher than that of Polyacrylonitrile fibre. FTIR revealed that the hydrolyzed Amidoxime fibres contained conjugated imine (-C=N-) sequences. Functionalization enhanced the sorption of amidoxime fibres by an increase of 20 % in the cationic exchange capacity. This was achieved by the part conversion of the cyano groups into the carboxylic acid groups. The fibres showed faster kinetics largely due the available exchange sites on the surface of the fibres hence the equilibration was achieved much quicker.
Nilsson, Lisa O. "Redesign of Alpha Class Glutathione Transferases to Study Their Catalytic Properties". Doctoral thesis, Uppsala : Acta Universitatis Upsaliensis : Univ.-bibl. [distributör], 2001. http://publications.uu.se/theses/91-554-5171-3/.
Texto completoMontaño, Inti Doraci Cavalcanti. "Otimização dinâmica do cultivo semi-contínuo de Pichia pastoris recombinante para produção das enzimas heterólogas alfa amilase e penicilina G acilase". Universidade Federal de São Carlos, 2010. https://repositorio.ufscar.br/handle/ufscar/4051.
Texto completoUniversidade Federal de Minas Gerais
This master's thesis project aims at studying the dynamic optimization of the operation of a bench scale (up to 5L) automated, agitated and aerated bioreactor, where the semi-continuous cultivation of recombinant Pichia pastoris is run. This yeast was cloned using the PGK1 promoter, which precludes the use of methanol as inducer, expressing constitutively the enzyme penicillin G Acylase (PGA) from Bacillus megaterium. While the group of molecular biology of DEQUFSCar is working on cloning the PGA, d P. pastoris expressing the enzyme - amylase from Bacillus subtilis was cultivated. This clone, provided by prof. Fernando Torres, UnB, uses the same construction and, therefore, its kinetics of growth and production should be very similar to the PGA s. Cultivation of recombinant Pichia pastoris was performed in flasks (skaker) using standard culture medium, aiming at obtaining kinetic data, which are the starting point for the escalation to a benchtop bioreactor. Following that, tests were performed in a 5L bioreactor in batch and fed batch operation modes. With the bioreactor data , kinetic parameters of growth, to be further used in the simulations, were estimated, using a hybrid algorithm (which combines the global method Simulated Annealing, with the local one Levenberg- Marquardt). This algorithm, is implemented in Matlab and available in the software library of Ladabio (Laboratory of Development and Automation of Bioprocesses ). From these data, models of microbial growth and of production were developed, following a classic approach (unstructured, non-segregated). Computer simulations using different feeding strategies and employing these models allowed mapping the dynamics of the system. From this information, optimal control strategies were proposed to define optimal feeding profiles. Cellular concentrations of 5.4 g/L (dry weight) were reached in shaker (20h of cultivation, when glucose is exhausted), expressing 218 U/mL of -amylase, compared to 11.4 g/L (dry weight) that were achieved in cultures in a bioreactor in batch simple (10h of cultivation, when glucose is exhausted), expressing 156 U/mL of -amylase In fed-batch cultures, cell concentrations of up to 45 g/L were achieved, expressing up to 260 U/mL of - amylase, with a productivity of 5.2 U/mL/ h. In fed-batch cultures of P. pastoris expressing PGA, cell concentrations of up to 35 g/L were achieved. Enzyme activity was not detected in the culture broth due to the effect of glycosylation. Immunodetection reaction confirmed the expression of the recombinant enzyme. Four specific growth rate equations were adjusted, with different types of inhibition by one product, detected at significant levels by liquid chromatography highperformance, but not yet identified. This metabolite was added as an inhibitor in kinetic models, using the peak areas, normalized as a pseudoconcentration. The best fit to the experimental data were the Monod kinetic model with non-competitive inhibition. Typical values obtained for the maximum specific growth and glucose/ cell conversion factor in bioreactor were max=0,24 h-1 and YX/S = 0,48. Algorithm for optimal control in open loop was developed and successfully implemented, providing a robust profiles of great power, whose validation is proposed as a continuation of this work.
Este mestrado se propoe a estudar a otimizacao dinamica de biorreator automatizado, tipo tanque agitado e aerado, em escala de bancada (ate 5L), onde se processa o cultivo semi-continuo de Pichia pastoris recombinante. Essa levedura foi clonada pelo grupo do prof. Fernando Torres, da UnB, utilizando o promotor PGK1, que dispensa a utilizacao de metanol como indutor, expressando constitutivamente a enzima -amilase de Bacillus subtilis. Durante a execucao deste mestrado, a enzima penicilina G acilase (PGA) de Bacillus megaterium esta sendo clonada pelo grupo de biologia molecular do DEQ-UFSCar usando a mesma construcao e, portanto, a cinetica de crescimento e producao da PGA heterologa devera ser muito semelhante as da -amilase, utilizada como estudo de caso para otimizacao do bioprocesso. Cultivos de Pichia pastoris recombinante foram realizados em frascos agitados, utilizando meio de cultivo padrao, objetivando o levantamento de dados cineticos, ponto de partida para o escalonamento em biorreator de bancada. Posteriormente, foram realizados ensaios em biorreator de 5L, em batelada e batelada alimentada. Com os dados obtidos nos cultivos em biorreator, e utilizando algoritmo hibrido para estimativa de parametros (que combina o metodo global Simulated Annealing, com o local de Levenberg-Marquardt), implementado em MatLab e disponivel no LaDABio (Laboratorio de Desenvolvimento e Automacao de Bioprocessos), foram ajustados parametros cineticos de crescimento, para serem utilizados nas simulacoes dos cultivos em biorreator. A partir dai, foi desenvolvido modelo de crescimento microbiano e de producao, utilizando um enfoque classico (modelo nao-estruturado, nao-segregado) para descrever o sistema. Com isso, torna-se possivel realizar simulacoes em computador usando diferentes estrategias de alimentacao, para mapear a dinamica do sistema. A seguir, foram desenvolvidos algoritmos de controle otimo em malha aberta para definicao de estrategias de alimentacao. Concentracoes celulares de 5,4 g/L (massa seca) foram alcancadas em cultivos em camara rotatoria (20h de cultivo, quando se esgota a glicose), expressando 218 U/mL de -amilase, comparado com 11,4 g/L(massa seca) que foram atingidos em cultivos em biorreator em bateladas simples (10h de cultivo, quando se esgota a glicose), expressando 156 U/mL de -amilase. Em cultivos em batelada alimentada concentracoes celulares de ate 45 g/L foram atingidas, expressando ate 260 U/mL de -amilase, com uma produtividade de 5,2 U/mL/h. Em cultivo em batelada alimentada de P. pastoris expressando PGA, concentracoes celulares de ate 35 g/L foram atingidas. Nao foi detectada atividade enzimatica no caldo de cultivo devido ao efeito da glicosilacao. Reacao de imunodeteccao confirmou a expressao da enzima recombinante. Foram ajustadas quatro equacoes de velocidade especifica de crescimento, com diferentes tipos de inibicao por um produto, detectado em niveis importantes por cromatografia liquida de alto desempenho, mas ainda nao identificado. Esse metabolito foi inserido como inibidor nos modelos cineticos, utilizando as areas dos picos, normalizadas, como uma pseudoconcentracao. Os melhores ajustes aos dados experimentais foram com modelo cinetico de Monod com inibicao nao-competitiva. Valores tipicos obtidos para a velocidade especifica maxima de crescimento e de fator de conversao glicose/celula em biorreator foram max = 0,24 h-1 e YX/S = 0,48. Algoritmo de controle otimo em malha aberta foi desenvolvido e implementado com sucesso, prevendo de forma robusta perfis otimos de alimentacao, cuja validacao fica proposta como continuidade deste trabalho.
Curien, Gilles. "Activation allostérique de la thréonine synthase d'A. Thaliana par la S-adénosylméthionine : mécanismes moléculaires et aspects régulateurs". Université Joseph Fourier (Grenoble ; 1971-2015), 1998. http://www.theses.fr/1998GRE10143.
Texto completoRabêlo, Flávio Henrique Silveira. "Sulfur supply as a sustainable technology for phytoextraction: its effects on cadmium uptake and detoxification in Panicum maximum Jacq. cv. Massai". Universidade de São Paulo, 2018. http://www.teses.usp.br/teses/disponiveis/64/64133/tde-07032019-134024/.
Texto completoA concentração de cádmio (Cd) no ambiente aumentou em décadas recentes, o que representa sério problema sócio-ambiental, visto que o Cd é tóxico para a maioria dos seres vivos. Por isso, é importante reduzir a concentração desse metal em ambientes contaminados e o uso de plantas adequadamente supridas com enxofre (S) pode contribuir para isso (fitoextração), uma vez que o S é componente de metabólitos envolvidos no combate ao estresse causado pelo Cd. Assim, o nosso objetivo com esse estudo foi avaliar o efeito do S na i) cinética de absorção do Cd, no ii) desenvolvimento radicular e na absorção de nutrientes, no iii) perfil metabólico e síntese de compostos tiol, e iv) na atividade do sistema antioxidante e fotossintético do capim-massai (Panicum maximum Jacq. cv. Massai), utilizado para fitoextração de Cd. Os estudos foram conduzidos em casa de vegetação utilizando-se vasos de 0,5 e 2,0 L para a condução do estudo de cinética de absorção de Cd (tratamentos representados pelas combinações das doses de S de 0,1 e 1,9 mmol L-1 e Cd de 0,1 e 0,5 mmol L-1) e para o estudo dos mecanismos de detoxificação de Cd (tratamentos representados pelas combinações das doses de S de 0,1; 1,9 e 3,7 mmol L-1 e Cd de 0,0; 0,1 e 0,5 mmol L-1), respectivamente. Os vasos foram distribuídos em blocos ao acaso, com quatro repetições. As plantas utilizadas no estudo de cinética foram expostas aos tratamentos durante 14 dias após as mesmas terem permanecido em soluções contendo apenas S durante 42 dias, enquanto as plantas utilizadas no estudo de detoxificação de Cd foram expostas aos tratamentos pelo período de 9 dias após terem crescido em soluções contendo apenas S por 44 dias. Ao final dos estudos, as plantas utilizadas foram colhidas e encaminhadas para análises. A velocidade máxima de absorção (Vmax) e o influxo apoplástico de Cd do capim-massai exposto à maior dose de Cd foram maiores quando as plantas foram supridas com a maior dose de S. O desenvolvimento radicular do capim-massai foi fortemente inibido quando as plantas foram expostas à dose de Cd de 0,5 mmol L-1, mas o adequado (1,9 mmol L-1) suprimento de S permitiu maior absorção de Cd, enquanto o suprimento excessivo (3,7 mmol L-1) de S diminuiu a formação de placas de ferro nas raízes das plantas. A lisina, cisteína, ornitina, arginina, triptofano e histidina foram acumulados em mais de um tecido nas plantas expostas ao Cd, assim como o galactinol. A glutationa (GSH), as fitoquelatinas (PCs) e seus homólogos foram fortemente induzidos pelo Cd, sendo que as plantas supridas com as doses de S de 1,9 e/ou 3,7 mmol L-1 apresentaram as maiores concentrações desses peptídeos. As plantas supridas com as maiores doses de S apresentaram menor peroxidação lipídica e maior taxa fotossintética, o que demonstra que o sistema antioxidante dessas plantas foi mais eficiente na atenuação dos danos causados pelo Cd. Diante desses resultados, fica claro que o capim-massai suprido adequadamente com S apresenta maior tolerância ao Cd, assim como maior potencial de fitoextração
Ravanel, Stéphane. "Biosynthèse de la méthionine chez les plantes supérieures : étude biochimique et moléculaire des enzymes de la voie de transsulfuration". Université Joseph Fourier (Grenoble ; 1971-2015), 1995. http://www.theses.fr/1995GRE10238.
Texto completoLencer, Doerthe Adriana. "Expression und Regulation von Zytokinrezeptor-mRNAs in Bronchialbiopsien von Patienten mit chronisch obstruktiver Lungenerkrankung /". 2005. http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&doc_number=014594483&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA.
Texto completoWang, Sihong. "Kinetics study of heat shock protein 70 expression". 2003. http://wwwlib.umi.com/cr/utexas/fullcit?p3116221.
Texto completoLin, Chiu-Fang y 林秋芳. "Differential Expression of mRNAs in Epithelia of Tubular Shell Glands of Brown Tsaiya Varying with Eggshell Strength". Thesis, 2001. http://ndltd.ncl.edu.tw/handle/00736888582993646061.
Texto completo國立臺灣大學
畜產學研究所
89
Deterioration of eggshell quality associated with many factors causes economic loss due to breakage during the egg transportation and processing. While many approaches including aspects related to nutrition, feeding management, or housing environment etc. have so far been employed years for enhancing the shell strength of chicken eggs, there are approximate 12% of all commercial eggshells broken or cracked between oviposition and consumption. The organic matrix, though it occupies only 1 ~ 2% of the whole eggshell contents, can be one of the most important components involved in eggshell quality. Therefore, in this present study, attempts were made to identify and characterize gene(s) differentially expressed 4.5 h post oviposition in epithelia of tubular shell-glands of brown Tsaiya and it is hopefully that the results will provide a better understanding to the molecular mechanisms related to eggshell formation. To achieve the said purpose, two herds of brown Tsaiya ducks showing significant difference in eggshell strength (ES), the high eggshell strength (HES) with an average ES value over 5.5 kg and the low eggshell strength (LES) with an average ES value less than 3.5 kg, were employed in the present study. Total RNA samples were extracted from epithelial cells of tubular shell gland 4.5 h post-ovipostion. Each RNA sample was subjected to analysis by the method of differential display reverse transcription polymerase chain reaction (DDRT-PCR) and two-step DDRT-PCR with a total of 72 primer sets designed. Comparisons were made to identify specific fragments of cDNA(s) between each sample amplified after the DDRT-PCR and two-step DDRT-PCR analysis. Among the 72 pairs of primer sets tested, only 8 primer-pairs were found to be effective in amplifying specific DNA fragments from the diversified products amplified. A total of 16 differential fragments of cDNA were obtained, including 11 fragments from the method of DDRT-PCR and 5 fragments from the two-step DDRT-PCR method. One of them, named TG36S, was further subjected to quantitative comparisons of its mRNA expressed between herds of HES- and LES-ducks, with the method of Real-Time Quantitive polymerase chain reaction (QPCR). Results appeared that the amount of TG36S mRNA expressed in the tubular shell glands from HES-ducks was 1.83 fold higher than that in the same tissue from LES-ducks. Moreover, to those HES-ducks, it was also noticed that much higher expression of TG36S, with some 7.57, 4.05, and 2.60 folds higher based on the amount of TG36S mRNA expressed, was found in epithelia of shell glands, tubular shell glands, and magnum, respectively, when comparisons were made to that of in epithelia of isthmus. Based on the fact that much higher expression of TG36S found in HES-ducks, particularly in epithelia of shell glands and tubular shell glands, it is worth to further investigate the physiological function of this gene product.
Hsiao, Yi-Jing y 蕭乂菁. "Study on the expression profiles of mRNAs and microRNAs in HLJ1-silenced non-small cell lung cancer cells". Thesis, 2008. http://ndltd.ncl.edu.tw/handle/48388871183461389865.
Texto completo國立中興大學
生物醫學研究所
96
Recently, lung cancer has dominantly increased and ranked up to the first place among Taiwanese women died from cancers. Therefore, understanding the tumorigenesis and metastasis in lung cancer are very important events. Previously study in our lab proved HLJ1 is a tumor suppressor. Moreover, HLJ1 expression was also associated with reduced cancer recurrence and improved overall survival in non-small cell lung cancer (NSCLC) patients. Over-expression of HLJ1 in cancer cells suppressed cell growth, invasion and migration. In order to clarify the role of HLJ1 in NSCLC cell, CL1-0, with the high HLJ1 expression but low capability of invasion and metastasis was utilized in this study. After reducing HLJ1 expression with RNA interference, the stable cell lines were successfully selected. It was observed that remolding of fiber actin induced reorganization of cell cytoskeleton, and the metastasis capability would increase significantly in these HLJ1-silenced cells. Furthermore, microarray was used to profile the expressions of genes and microRNAs in HLJ1-silenced cells and analyzed the results by composite regulatory signature database which was developed in our lab. The results revealed that HLJ1 could not only regulate the genes involved in cytoskeleton and cell cycle, but also moderate the signaling pathways such as MAPK, Wnt and epithelial-to-mesenchymal transition (EMT) pathway. In cytoskeleton signaling pathway, Rac1-GTP was prove to be up-regulated in HLJ1-silenced clones by GST pull-down assay. In these pathways, 43 genes were selected for further validation of gene expression patterns by real-time PCR. There are 34 genes with matched expression patterns analyzed by chip and real-time PCR, such as NTS、cyclin D1 and ROCK1 etc. Moreover, there are 15 genes as transcription factors, including PITX2、HOXA1、MEIS2、HOXA5、FOXC1、CITED2、PAX9 and E2F5 those were up-regulated and ATF3、KLF5、ATBF1、HEY1、HOXB5、S100A10 and TAF5 were down-regulated in HLJ1-silenced cells. For example, transcription factor PITX2 and HOXA1 would up-regulate the oncogene cyclin D1. Aadditionally, profiling of the microRNA expressions in HLJ1-silenced cells by microRNA chip was no significantly different with HLJ1-non-silenced cells. (From 0.6 to 1.6 fold) Furthermore, the nuclear aggregates of β-catenin were found in HLJ1-silenced cells. The function of β-catenin in nucleus was proved previously to activate transcription. Those evidences show that HLJ1 may regulate β-catenin localization to activate the transcription of the downstream genes, including PITX2 and cyclin D1 genes. In conclution, HLJ1 mediates the activation of Rac1 and the localization of β-catenin to regulate the expression of downstream genes, and moderate the cell morphology and the abilities of invasion and migration in cancer cells. These studies give a new annotation of HLJ1 in cancer metastasis and invasion mechanism. But in the other signaling pathways, the role of HLJ1 still needs more efforts to research. The combined microarray and bioinformatic anyalsis provides more directions for further investigation. In the future, it is expected that can facilitate on the treatment of cancer.
Lin, Fang-Yin y 林芳因. "IL-1 may up-regulate the expression of IL-6 and ColIA1 mRNAs in the cells of dihydropyridine induced gingival". Thesis, 2009. http://ndltd.ncl.edu.tw/handle/42883920578723368475.
Texto completo臺北醫學大學
牙醫學系碩博士班
97
Nifedipine is a well known calcium channel blockers for treatment of hypertension and angina. The side effect as nifedipine-induced gingival overgrowth (NIGO) can be frequently found in clinic. NIGO cells, as stimulated by IL-1β, may elicit a stronger IL-6 expression and collagen production. Moreover, Akt phosphorylation was correlated to collagen overproduction. We propose a hypothesis that PI3K/Akt/NFκB pathway is one of the possible mechanisms which may be related to the over expression of IL-6 and overproduction of extra-cellular collagen in dihydropyridine induced gingival overgrowth (DIGO) cells. In this study, we will use western blot technique to detect the expression of Akt. Moreover, real-time PCR is utilized to explore the expression of IL-6 and collagen-I mRNA. Sircol dye assay was used to detect total collagen concentration. Finally, we will interpret the correlation of p-Akt/Akt, IL-6 and collagen by non-parametric Spearman correlation coefficient. Result revealed that by stimulating with IL-1β, the value of p-Akt/Akt ratio, the expression of IL-6 and collagen-I mRNA, and the concentration of IL-6 and collagen in supernatant were all increased. In addition to the expression of IL-6 mRNA, the other three factor described above were higher in DIGO group then in healthy group stimulated by IL-1β. Spearman correlation coefficient shows high relationship between the value of p-Akt/Akt, the expression of IL-6 and collagen-I mRNA, and the concentration of IL-6 in supernatant. The data mentioned above approve that Akt is in connection with gingival overgrowth.