Este trabalho apresenta a tradução dos fragmentos supérstites do poema épico Anais de Quinto Ênio (239 ca. 169 a. C.). Uma seção introdutória trata de discutir o que se sabe sobre o poeta, partindo das citações dos autores antigos que a ele se referiram. Faz ainda parte dessa seção inicial um estudo sobre os Anais, observando o poema segundo as visões dos antigos e também de acordo com o que é hoje, na forma fragmentária em que chegou a nós, cujo elemento unificador se centra no trabalho filológico de críticos de todo o mundo. O cerne do trabalho consiste na tradução e anotação dos 420 fragmentos tomados à edição italiana de Valmaggi (1945). Os comentários se baseiam sobretudo nas reflexões de Skutsch (1985), Steuart (1976), Warmington (1988) e Vahlen (1967), partindo da contextualização de cada fragmento, assinalando o tema a que esteja ligado e, por conseguinte, explicando por que tal fragmento foi incluído no canto de que faz parte. Em seguida, ocupamo-nos de analisar o fragmento, ressaltando motivações estilísticas e empregos lingüísticos, em busca do significado do texto-fragmento, o que se faz com o uso de recursos diversos, como o auxílio de diferentes dicionários, a comparação da mesma palavra em distintos fragmentos dos Anais ou de outras obras enianas, ou ainda o estudo do emprego de uma palavra em contextos semelhantes de outros autores, ou em diferentes contextos de autores contemporâneos de Ênio. / This thesis presents the translation of the remaining fragments of the epic poem Annals by Quintus Ennius (239ca. 169 BC). An introductory section discusses what is known about the poet, taking the ancient authors quotes that refer to him as a starting-point. In this initial section a study on the Annals is also included; it observes the poem according to the ancients point of view and to what it is today, in the fragmentary form in which it has come down to us, the philological work of critics from all over the world being its unifying element. The kernel of this text consists of the translation and commentary of the 420 fragments taken from the Italian edition of Valmaggi (1945). The comments are based primarily on the contributions of Skutsch (1985), Steuart (1876), Warmington (1988) and Vahlen (1967), and the contextualization given to each fragment, since such procedure aids and enables my search for the theme it is connected with and thus explains why each fragment was included in the book it is part of. I then analize the fragments one by one, mark its stylistic motivations and linguistic uses, and search for the meaning of each text-fragment, which is done through diverse resources, such as the help of different dictionaries, word comparison in distinct fragments either of the Annals or other Ennius works, and the study of a words usage in similar contexts found in other authors, and in different contexts of Ennius contemporary authors.

Essa dissertação é o resultado dos questionamentos e das inquietações suscitadas a partir do encontro das narrativas de ficções científicas e das ideias feministas. Esses pulsares teóricos transpassam em alguns momentos minha trajetória acadêmica, tentando, concomitantemente, conciliar e incorporar essas duas formas literárias. A discussão basilar advém da leitura de The Female Man, de Joanna Russ, em um encadeamento sutil com a minha perspectiva enquanto sujeito mulher em contato com esses universos. Por fim, busca-se desvendar os fragmentos narrativos que estão impregnados de poderosas imagens subversivas e palavras transgressoras que convocam as leitoras e leitores a repensar, resistir e reformular a realidade ao seu redor. / This thesis is the result of questioning and restlessness raised by the encounter of science fiction narratives and feminist ideas. These theoretical pulses trespass at some moments my academic trajectory, trying concomitantly to harmonize and to embody these two literary types. The underlying discussion comes from the reading of The Female Man, by Joanna Russ, inside a sutil chaining with my perspective as woman subject in contact with these universes. Lastly, It seeks to unravel the narrative fragments that are impregnated with powerful subversive images and transgressive words that invite male and female readers to rethink, resist and reshape the reality around them

Lo-Fi vem de Low Fidelity, que em inglês significa baixa fidelidade. Na música, se transformou em linguagem sonora a partir da década de 1970, quando alguns artistas, buscando soluções econômicas para a autoprodução e independência em relação às gravadoras, passaram a utilizar gravadores caseiros para fazer seus próprios registros. A partir desse ato de libertação e experimentação, se obteve, por transbordamento, uma sonoridade ruidosa, crua, de baixa fidelidade de gravação. Mas, ao mesmo tempo, o ruído produzido por essa produção marginal engendrou cruzamentos autênticos e espontâneos que logo formaram uma paisagem sonora de grande vitalidade. De forma contrária, o Hi-fi, ou High-Fidelity, busca na alta-fidelidade a idealização e a perfeição do seu produto final que, uma vez ligado à indústria fonográfica e à alta tecnologia, tende a ser direcionado pelas lógicas do mercado. Pensado e produzido como objeto-fim, o fazer arquitetural contemporâneo vem se aproximando da gravação hi-fi: delimitado, ordenado por estratos, classificações e tipos que a definem como objeto-produto. Nessa lógica, a arquitetura torna-se imagem, espetáculo pronto para o consumo e descarte. Propõe-se, neste trabalho, uma inflexão, um deslocamento do pensar arquitetural a partir da aproximação metafórica acionada pelo modo lo-fi . / Lo-Fi is the acronym that stands for Low Fidelity. In the music industry, during the70’s it became a trendy language when artists, searching economical solutions for self-production and independence from record companies, using homemade devices for sound recording. Following this first experiment of liberation, was obtained a noisy overflowing sonority, raw, or as it became known, low fidelity of the recording. However, at the same time the noise produced by this marginal production created an authentic and spontaneous blend that soon formed a vivid sound scene. Contrariwise the Hi-fi, or High Fidelity, seeks the idealization of perfection as its final product which, once merged to the phonographic industry and the high technology, tends to be driven by the logic of the market. Thought and produced as a final product, contemporary architecture follows the steps of the hi-fi recording: delimited, ordered in layers, classifications and by stereotypes that define it as an object-product. In this logic, architecture becomes an image, a spectacle ready for consumption and disposal. This work proposes an inflexion, a displacement on the architectural thinking from the metaphorical approach triggered by the lo-fi mode. Flaws by noise, by experimentation, by the potency of invention leads to a turning point in the making process, broader, returning to the idea of the unfinished, unexpectedly, guiding into a process that its meaning is not in the final product, but rather in the path itself, in the desire for transformation, in the unknown. Reflecting about the process as lo-fi music evokes is returning to the initial sketch, it is decomposing and questioning its nature, it is exposing the traces. It is bringing back the architecture in the sense of discovery, as a body in experimentation, as the surprise that reveals the unforeseen crossing of distinct times. The intention here is, based from speculative narratives, to try the deconstruct of some consolidated practices on the design process catalyzed by the music action. Pointing analogous approximations and representations of other architectural assemblies that are not represented by the narratives determined and exhausted by the logic of the market.

La glycoprotéine VI (GP VI) des plaquettes sanguines humaines est le récepteur principal du collagène, composé le plus thrombogénique d'une paroi vasculaire lésée. Ainsi, GPVI est souvent considérée comme une cible de premier plan pour développer des tests diagnostiques ou des stratégies thérapeutiques innovantes, efficaces et sûres afin d'améliorer encore la prise en charge des accidents ischémiques. Les anticorps monoclonaux et leurs fragments actifs produits par ingénierie moléculaire constituent aujourd'hui une nouvelle classe de biomolécules en plein essor avec des propriétés bien adaptées à des applications thérapeutiques et diagnostiques. Notre groupe a produit plusieurs anticorps monoclonaux anti-GPVI par immunisation génique de souris. Ces anticorps ont une affinité élevé pour leur cible. Ils de distinguent les uns des autres par leur spécificité épitopique ainsi que par les effets engendrés par leur liaison à GPVI. Parmi ces anticorps, l'un présente un fort potentiel diagnostique parce qu'il reconnait les formes mono- et dimériques de GPVI, mais sa liaison aux plaquettes peut induire une activation ou la perte de GPVI. Un autre anticorps présente un fort potentiel thérapeutique parce que ses fragments actifs monovalents obtenus par papaïnolyse neutralisent l'interaction entre les plaquettes et le collagène, sans activer les plaquette. Cependant, l'origine xénogénique de cet anticorps est responsable d'une forte immunogénicité qui en interdit des applications en médecine humaine. Dans cette étude, nous avons conçus un fragments variable d'anticorps simple chaine (scFv) utile pour quantifier l'expression de la GPVI à la surface des plaquettes sanguines. Ce scFv a été reformaté de façon à lui insérer un motif de reconnaissance de la Protéine L (PpL) qui facilite sa détection et sa purification sans avoir recours à un peptide "drapeau". Nous avons également humanisé et créé plusieurs fragments d'anticorps recombinants monovalents inhibiteurs de l'interaction GPVI / collagène. Ces fragments d'anticorps présentent un potentiel thérapeutique élevé. / Human platelets glycoprotein VI (GPVI) is evidenced to be a platelet receptor of major importance in the occurrence of arterial thrombosis. Thus, it can be considered to be of great interest in diagnosis and therapeutic of atheriosclerotic diseases. Antibodies are powerful molecules which can be used in both diagnostic as well as for therapeutic purposes due to their unique characteristics. Monoclonal and recombinant antibodies have antigen restricted specificity, high affinity and can be used in various assays. Moreover, the good knowledge of their structure and molecular engineering facilities now allows the antibody modulation according to desired properties.Our group has already produced several monoclonal antibodies to human GPVI by gene gun immunization against the immunoadhesin hGPVI-Fc, which differ in fine epitopespecificity, affinity and other functional properties (Lecut et al. 2003). One, 3J24, with diagnostic potential while the other, 9O12, has a therapeutic potential because it blocks the binding of GPVI to collagen. Its Fab fragment has been extensively characterized in vitro,ex vivo and in vivo for its antithrombotic properties.Here, we designed and reshaped a single-chain antibody fragment (scFv) based on 3J24variable domains for the quantification of GPVI with diagnostic potential. We were also involved in the design, production and functional evaluation of humanized anti-GPVI recombinant antibody fragments (scFvs and Fabs) with therapeutic properties.

Este trabalho apresenta a tradução dos fragmentos supérstites do poema épico Anais de Quinto Ênio (239 ca. 169 a. C.). Uma seção introdutória trata de discutir o que se sabe sobre o poeta, partindo das citações dos autores antigos que a ele se referiram. Faz ainda parte dessa seção inicial um estudo sobre os Anais, observando o poema segundo as visões dos antigos e também de acordo com o que é hoje, na forma fragmentária em que chegou a nós, cujo elemento unificador se centra no trabalho filológico de críticos de todo o mundo. O cerne do trabalho consiste na tradução e anotação dos 420 fragmentos tomados à edição italiana de Valmaggi (1945). Os comentários se baseiam sobretudo nas reflexões de Skutsch (1985), Steuart (1976), Warmington (1988) e Vahlen (1967), partindo da contextualização de cada fragmento, assinalando o tema a que esteja ligado e, por conseguinte, explicando por que tal fragmento foi incluído no canto de que faz parte. Em seguida, ocupamo-nos de analisar o fragmento, ressaltando motivações estilísticas e empregos lingüísticos, em busca do significado do texto-fragmento, o que se faz com o uso de recursos diversos, como o auxílio de diferentes dicionários, a comparação da mesma palavra em distintos fragmentos dos Anais ou de outras obras enianas, ou ainda o estudo do emprego de uma palavra em contextos semelhantes de outros autores, ou em diferentes contextos de autores contemporâneos de Ênio. / This thesis presents the translation of the remaining fragments of the epic poem Annals by Quintus Ennius (239ca. 169 BC). An introductory section discusses what is known about the poet, taking the ancient authors quotes that refer to him as a starting-point. In this initial section a study on the Annals is also included; it observes the poem according to the ancients point of view and to what it is today, in the fragmentary form in which it has come down to us, the philological work of critics from all over the world being its unifying element. The kernel of this text consists of the translation and commentary of the 420 fragments taken from the Italian edition of Valmaggi (1945). The comments are based primarily on the contributions of Skutsch (1985), Steuart (1876), Warmington (1988) and Vahlen (1967), and the contextualization given to each fragment, since such procedure aids and enables my search for the theme it is connected with and thus explains why each fragment was included in the book it is part of. I then analize the fragments one by one, mark its stylistic motivations and linguistic uses, and search for the meaning of each text-fragment, which is done through diverse resources, such as the help of different dictionaries, word comparison in distinct fragments either of the Annals or other Ennius works, and the study of a words usage in similar contexts found in other authors, and in different contexts of Ennius contemporary authors.

Made available in DSpace on 2017-07-10T17:30:28Z (GMT). No. of bitstreams: 1 Daniele Moraes.pdf: 8273735 bytes, checksum: 9f8283d1a437ef50a7fc91af11f5445a (MD5) Previous issue date: 2014-04-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This research aimed to identify the procedures required for nature trails in forested areas´ deployment and management, considering benefits and problems of its use in small farms. Through literature review and empirical studies, it was possible to survey the methods and techniques of deployment, use and management of trails in forested areas, and to evaluate physical and environmental conditions, the user profile and interpretative activities in two ecological trails located in a small farm open to visitors, called Recanto Renascer, located in the municipality of Francisco Beltrão, state of Parana, Brazil. To assess the impact on the trails, the methodology Monitoring the Impact of the Visitation (IVM) was used. For the analysis of the attractiveness of each track, we used the methodology of Interpretative Attractiveness Index Points (IAPI). Soil analysis for verification of soil compaction on the trails were also done. The results obtained from the research used to develop proposals for physical- environmental and interpretive adequacy of the two tracks of Recanto Renascer, beyond the preparation of a material with technical information to owners of farms interested in establishing nature trails or in adapting existing trails. / Esta pesquisa teve como objetivo principal conhecer os procedimentos necessários para a implantação e manejo de trilhas ecológicas em áreas florestais, considerando benefícios e problemáticas de seu uso em pequenos estabelecimentos rurais. Através da revisão bibliográfica e dos estudos empíricos realizados, foi possível fazer um levantamento dos métodos e técnicas de implantação, uso e manejo das trilhas em áreas florestais, além de avaliar as condições físico-ambientais, o perfil dos usuários e as atividades interpretativas desenvolvidas em duas trilhas ecológicas localizadas em um pequeno estabelecimento rural aberto à visitação, denominado Recanto Renascer, no município de Francisco Beltrão-PR. Para o levantamento dos impactos nas trilhas, foi utilizada a metodologia Monitoramento do Impacto da Visitação (MIV). Para a análise dos atrativos de cada trilha, utilizou-se a metodologia Índice de Atratividade de pontos Interpretativos (IAPI). Também foram realizadas análises de solo, para verificação da compactação dos solos nas trilhas. Os resultados obtidos com a pesquisa permitiram desenvolver propostas de adequação físico-ambiental e interpretativa das duas trilhas do Recanto Renascer, além da elaboração de um material técnico-informativo destinado a proprietários de estabelecimentos rurais interessados em implantar trilhas ecológicas ou adequar as já existentes.

The rate of DNA mutation is known to fluctuate across the genome but the patterns of mutation rate variation and molecular causes are poorly defined. It is important to understand these patterns of mutation as they influence where deleterious mutations are likely to arise and how rapidly sequences are likely to accumulate change between species, a measure often used as a proxy for functional constraint. In this work I investigate the relationship between DNA replication and apparent mutation hotspots adjacent to transcription factor binding sites. In eukaryotes both DNA strands are replicated simultaneously, the leading strand as a continuous stretch and the lagging strand as a series of discrete Okazaki fragments that are subsequently ligated together. Some transcription factors are able to bind the DNA lagging strand during replication and act as a partial barrier to DNA polymerase, resulting in the accumulation of Okazaki fragment junctions adjacent to these sites. I find that mutation rate is correlated genome wide with Okazaki junction frequency, suggesting that Okazaki junction processing may be error-prone. We present a mechanistic hypothesis to explain this locally elevated mutation rate and propose a role for lagging strand replication and its error-prone Pol α tract retention in the formation of these hotspots. I test this hypothesis using Okazaki fragment sequencing data from the yeast Saccharomyces cerevisiae to identify peaks in Okazaki junctions. When these peaks are aligned and orientated, so that the direction of lagging strand replication is uniform, I find a peak in substitution rate immediately downstream of Okazaki junctions, precisely where Pol α tract retention is predicted to occur. Novel binding motifs are identified within the underlying DNA of these junctions that can be assigned to known strong and fast-binding transcription factors, previously implicated in the phasing of nucleosomes, such as Reb1. I show that mutation hotspots adjacent to transcription factor binding sites are a conserved feature of eukaryotic genomes. In the human genome I predict sites of preferential Pol α retention using DNase I hypersensitivity footprint data. We observe that those footprints predicted as germline-specific manifest an elevated mutation signature. I propose that the rapid binding of some transcription factors to DNA following replication is required for nucleosome positioning or other important functions, however this incurs a cost in terms of locally elevated mutation rate adjacent to and within the sequence specific binding site. As a consequence these binding sites are biologically important mutational hotspots whose functional significance has been systematically underestimated by standard measures of sequence constraint.

Made available in DSpace on 2017-05-12T14:42:04Z (GMT). No. of bitstreams: 1 Daniele Moraes.pdf: 8273735 bytes, checksum: 9f8283d1a437ef50a7fc91af11f5445a (MD5) Previous issue date: 2014-04-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This research aimed to identify the procedures required for nature trails in forested areas´ deployment and management, considering benefits and problems of its use in small farms. Through literature review and empirical studies, it was possible to survey the methods and techniques of deployment, use and management of trails in forested areas, and to evaluate physical and environmental conditions, the user profile and interpretative activities in two ecological trails located in a small farm open to visitors, called Recanto Renascer, located in the municipality of Francisco Beltrão, state of Parana, Brazil. To assess the impact on the trails, the methodology Monitoring the Impact of the Visitation (IVM) was used. For the analysis of the attractiveness of each track, we used the methodology of Interpretative Attractiveness Index Points (IAPI). Soil analysis for verification of soil compaction on the trails were also done. The results obtained from the research used to develop proposals for physical- environmental and interpretive adequacy of the two tracks of Recanto Renascer, beyond the preparation of a material with technical information to owners of farms interested in establishing nature trails or in adapting existing trails. / Esta pesquisa teve como objetivo principal conhecer os procedimentos necessários para a implantação e manejo de trilhas ecológicas em áreas florestais, considerando benefícios e problemáticas de seu uso em pequenos estabelecimentos rurais. Através da revisão bibliográfica e dos estudos empíricos realizados, foi possível fazer um levantamento dos métodos e técnicas de implantação, uso e manejo das trilhas em áreas florestais, além de avaliar as condições físico-ambientais, o perfil dos usuários e as atividades interpretativas desenvolvidas em duas trilhas ecológicas localizadas em um pequeno estabelecimento rural aberto à visitação, denominado Recanto Renascer, no município de Francisco Beltrão-PR. Para o levantamento dos impactos nas trilhas, foi utilizada a metodologia Monitoramento do Impacto da Visitação (MIV). Para a análise dos atrativos de cada trilha, utilizou-se a metodologia Índice de Atratividade de pontos Interpretativos (IAPI). Também foram realizadas análises de solo, para verificação da compactação dos solos nas trilhas. Os resultados obtidos com a pesquisa permitiram desenvolver propostas de adequação físico-ambiental e interpretativa das duas trilhas do Recanto Renascer, além da elaboração de um material técnico-informativo destinado a proprietários de estabelecimentos rurais interessados em implantar trilhas ecológicas ou adequar as já existentes.

A tuberculose (TB) é considerada uma das principais doenças infecciosas e apresenta fatores críticos como a relação com o HIV/AIDS, tratamento longo e a resistência a múltiplos fármacos. A enzima di-hidrofolato redutase das micobactérias (mtDHFR) é um alvo pouco explorado e apresenta grande potencial para o desenvolvimento de novos fármacos contra TB. Estudos preliminares obtiveram fragmentos com baixa afinidade à mtDHFR, entretanto com potencial para otimização. Com isso, o fragmento foi usado como protótipo para a proposição de 22 análogos. Os compostos foram planejados utilizando informações sobre ligantes e a estrutura tridimensional de mtDHFR, além do biososterismo como estratégia norteadora. Os ensaios de docking molecular com a mtDHFR revelaram que os análogos propostos tiveram escores interessantes e, além disso, a inserção de substituintes demonstrou favorecer a ligação à enzima, o que corroborou o planejamento. Com isso, sintetizou-se 22 análogos planejados e o protótipo MB872, por meio de protocolos de alquilação, hidrólise e cicloadição 1,3 dipolar para os compostos com anéis triazol e tetrazol. Os compostos foram obtidos com rendimentos de bom a ótimo (60 ~ 90%) e suas estruturas foram elucidadas por RMN 1H e 13C. Os resultados do ensaio de inibição enzimática corroboraram com os dados de docking, uma vez que a presença do grupo carboxílico revelou ser importante para a atividade. Além disso, alguns dos compostos revelaram atividades interessantes, entre 8 a 40 µM, sendo que o mais ativo apresentou IC50 de 7 µM. Ensaios de cinética enzimática com o análogo mais ativo indicou uma inibição não competitiva com o substrato natural da enzima, uma vez que os valores de Km se mantiveram constantes, enquanto Vmax decaiu (0,22 µM e 0,43 - 0,34 ΔFU/min, respectivamente). Os análogos sintetizados foram mandados para ensaio in vitro para avaliar a atividade frente a micobactéria. / Tuberculosis (TB) is an important infectious disease and presents critical factors such as the relationship with HIV / AIDS, long treatment and resistance to multiple drugs. The enzyme dihydrofolate reductase from mycobacteria (mtDHFR) is a poorly explored and presents great potential to be a target for new drugs against TB. Preliminary studies have obtained fragments with low affinity to mtDHFR, but with potential to become lead compounds. Therefore, the fragment was used as a prototype for 22 analogues proposed in this work. The compounds were designed using bioisosterism, information about ligands and the three-dimensional structure of mtDHFR. Molecular docking assays with mtDHFR revealed satisfactory scores for anlogues. Furthermore, the insertion of substituents seemed to increase the affinity with the enzyme. Thereby, twenty two analogues and prototype were synthesized using alkylation, hydrolysiss and 1,3-dipolar cycloaddition methods. The compounds were obtained in good yields (60 ~ 90%) and their structures were elucidated with 1H and 13C NMR spectroscopy. The enzymatic affinity assay corroborates docking results, because the presence of carboxyl group showed to be important for the activity. Furthermore, some of the compounds revealead interesting activities, ranging 8 to 40 µM. The most active showed IC50 of 7 µM and enzyme kinetics assays indicated noncompetitive inhibition with natural enzyme substrate. The synthesized analogs were sent for in vitro assay to assess mycobacteria activity.

Tam, Hou Si. / 33 in title is subscript. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 174-187). / Abstracts in English and Chinese. / THESIS COMMITTEE --- p.i / ACKNOWLEDGEMENTS --- p.ii / ABSTRACT --- p.iii / 摘要 --- p.v / TABLE OF CONTENTS --- p.vii / LIST OF FIGURES --- p.xii / LIST OF TABLES --- p.xvii / LIST OF ABBREVIATIONS --- p.xviii / CHAPTER / Chapter 1. --- INTRODUCTION / Chapter 1.1 --- Malaria --- p.1 / Chapter 1.2 --- Malaria is a public health problem --- p.1 / Chapter 1.3 --- Malarial parasite --- p.3 / Chapter 1.4 --- Life cycle of P. falciparum --- p.3 / Chapter 1.4.1 --- The pre-erythrocytic stage --- p.3 / Chapter 1.4.2 --- The asexual erythrocytic stage --- p.3 / Chapter 1.4.3 --- The sexual transmission stage --- p.6 / Chapter 1.5 --- Chemoprophylaxis and chemotherapy of malaria --- p.7 / Chapter 1.6 --- Drug resistance of malaria parasite --- p.7 / Chapter 1.7 --- The progress for malaria vaccine --- p.10 / Chapter 1.8 --- Vaccine candidates for asexual erythrocytic stage --- p.11 / Chapter 1.9 --- Merozoite Surface Protein-1 (MSP-1) --- p.13 / Chapter 1.9.1 --- Structure of MSP-1 --- p.13 / Chapter 1.9.2 --- The processing of MSP-1 --- p.17 / Chapter 1.9.3 --- MSP-1 as a blood-stage vaccine --- p.19 / Chapter 1.9.4 --- The vaccine potency of MSP-133 --- p.23 / Chapter 1.10 --- Merits of E. coli expression system --- p.25 / Chapter 1.11 --- Aim of study --- p.26 / Chapter 2. --- MATERIALS AND METHODS / Chapter 2.1 --- Materials --- p.30 / Chapter 2.2 --- Methods --- p.39 / Chapter 3. --- EXPRESSION AND PURIFICATION OF RECOMBINANT MSP-l33kv+19 PROTEIN / Chapter 3.1 --- Introduction --- p.63 / Chapter 3.2 --- Results / Chapter 3.2.1 --- Construction of pET32a/MSP-l33kv+19 expression vector --- p.64 / Chapter 3.2.2 --- SDS-PAGE analysis of the expressed protein --- p.74 / Chapter 3.2.3 --- Western blot analysis of the expressed protein --- p.78 / Chapter 3.2.4 --- Modification of the expression conditions --- p.78 / Chapter 3.2.5 --- Protein purification by IMAC --- p.82 / Chapter 3.2.6 --- Cleavage of fusion partner from the rMSP-133kv+19 protein --- p.82 / Chapter 3.2.7 --- Verification of non-fused recombinant MSPl33kv+19 protein by N-terminal amino acid sequencing --- p.86 / Chapter 3.2.8 --- Separation of target protein from the fusion mixture by IMAC --- p.86 / Chapter 3.2.9 --- Separation of digestion product by Size Exclusion Chromatography --- p.89 / Chapter 3.2.10 --- Conformational test of the purified protein --- p.89 / Chapter 3.2.11 --- Separation of target protein from contaminants by Anion-Exchange Chromatography --- p.92 / Chapter 3.2.12 --- Separation of target protein from contaminants by Immuno-Affinity Chromatography --- p.95 / Chapter 3.3 --- Conclusion --- p.95 / Chapter 4. --- IMMUNOLOGICAL CHARACTERIZATION OF BACTERIAL EXPRESSED rMSP-l33kv+19 / Chapter 4.1 --- Introduction --- p.97 / Chapter 4.2 --- Results / Chapter 4.2.1 --- Immunogenicity of recombinant NfMSP-133kV+19 protein --- p.98 / Chapter 4.2.2 --- Specificity of anti-NfMSP-133kv+19 sera to MSP-l33kv. MSP-l33 and MSP-l19 --- p.98 / Chapter 4.2.3 --- Cross reactivity of anti-MSP-133kv+19 and anti-BVp42 serum --- p.103 / Chapter 4.2.4 --- Competitive ELISA --- p.103 / Chapter 4.2.5 --- Test for the presence of inhibitory B-cell epitopes on rMSP-l33kv+19 --- p.111 / Chapter 4.2.6 --- In vitro parasitic growth inhibition assay --- p.113 / Chapter 4.3 --- Conclusion --- p.115 / Chapter 5. --- EXPRESSION AND PURIFICATION OF RECOMBINANT MSP-l33kc+19 PROTEIN / Chapter 5.1 --- Introduction --- p.116 / Chapter 5.2 --- Results / Chapter 5.2.1 --- Construction of pET32a/MSP-133kv+19 expression vector --- p.117 / Chapter 5.2.2 --- Expression of recombinant MSP-133kc+19 protien (rMSP-133kc+19) --- p.124 / Chapter 5.2.3 --- Purification of rMSP-l33kc+19 by IMAC --- p.127 / Chapter 5.2.4 --- Cleavage of fusion partner from target protein --- p.127 / Chapter 5.2.5 --- Construction of pRSETA/MSP-l3X33kc+19 expression vector --- p.135 / Chapter 5.2.6 --- SDS-PAGE analysis of the protein expression --- p.146 / Chapter 5.3 --- Conclusion --- p.153 / Chapter 6. --- DISCUSSION / Chapter 6.1 --- Expression of rMSP-l33kv+19 --- p.154 / Chapter 6.2 --- Purification of rMSP-l3.3kv+19 --- p.156 / Chapter 6.3 --- Conformational test of rMSP-133kv+19 --- p.157 / Chapter 6.4 --- Biological and immunological activity of NfMSP-133kv+19 --- p.158 / Chapter 6.5 --- Expression of rMSP-133kc+19 --- p.166 / Chapter 6.6 --- Future prospects --- p.167 / REFERENCES --- p.174 / APPENDICES / Chapter 1. --- HiTrap NHS-activated HP for ligand coupling procedure --- p.188 / Chapter 2. --- Reuse of Ni+-NTA Resin procedure --- p.190 / Chapter 3. --- Sequence alignment of MSP-133 (MAD20 & Welcome/Kl alleles) --- p.191 / Chapter 4. --- Nucleotide sequence and amino acid sequence of P. falciparum MSP-l33kv+19 --- p.192 / Chapter 5. --- Nucleotide sequence and amino acid sequence of P. falciparum MSP-l33kc+19 --- p.193 / Chapter 6. --- "Nucleotide sequence and amino acid sequence of P. falciparum MSP-142 (3D7 isolate, MAD20 allele)" --- p.194 / Chapter 7. --- Amino acid sequence of Plasmodium falciparum MSP-l42 --- p.195