158 resultados para Construção de cassetes de superexpressão

em Universidade Federal do Rio Grande do Norte(UFRN)


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Flowering is a fundamental process in the life cycle for plant. This process is marked by vegetative to reproductive apical meristem conversion, due to interactions between several factors, both internal and external to plant. Therefore, eight subtractive libraries were constructed using apical meristem induced or not induced for two contrasting species: Solanum lycopersicum cv. Micro-Tom and Solanum pimpinellifolium. Several cDNAs were identified and among these, were selected two cDNAs: one homologous cDNA to cyclophilin (LeCYP1) and the other to Auxin repressed protein (ARP). It has observed that LeCYP1 and ARP genes are important in the developmental process to plants. In silico analysis, were used several databases with the exclusion criterion E-value <1.0x10-15. As a result, conservation was observed for proteins analyzed by means of multiple alignments and the presence of functional domains. Then, overexpression cassettes were constructed for the ARP cDNA in sense and antisense orientations. For this step, it was used the CaMV35S promoter. The cDNA orientation (sense or antisense) in relation to the promoter was determined by restriction enzymes and sequencing. Then, this cassette was transferred to binary vector pZP211 and these cassettes were transferred into Agrobacterium tumefaciens LBA4404. S. lycopersicum cv. Micro-Tom (MT) and MT-Rg1 plants were transformed. In addition, seedlings were subjected to hormone treatments using a synthetic auxin (- naphthalene acetic acid) and cyclosporin A (cyclophilin inhibitor) treatments and it was found that the hormone treatment there were changes in development of lateral roots pattern, probably related to decreases in auxin signaling caused by reduction of LeCYP1 in MT-dgt plants while cyclosporin A treatments, there was a slight delay in flowering in cv. MT plants. Furthermore, assay with real-time PCR (RT-qPCR) were done for expression level analysis from LeCYP1 and ARP in order to functionally characterize these sequences in tomato plants.

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Flowering is a fundamental process in the life cycle for plant. This process is marked by vegetative to reproductive apical meristem conversion, due to interactions between several factors, both internal and external to plant. Therefore, eight subtractive libraries were constructed using apical meristem induced or not induced for two contrasting species: Solanum lycopersicum cv. Micro-Tom and Solanum pimpinellifolium. Several cDNAs were identified and among these, were selected two cDNAs: one homologous cDNA to cyclophilin (LeCYP1) and the other to Auxin repressed protein (ARP). It has observed that LeCYP1 and ARP genes are important in the developmental process to plants. In silico analysis, were used several databases with the exclusion criterion E-value <1.0x10-15. As a result, conservation was observed for proteins analyzed by means of multiple alignments and the presence of functional domains. Then, overexpression cassettes were constructed for the ARP cDNA in sense and antisense orientations. For this step, it was used the CaMV35S promoter. The cDNA orientation (sense or antisense) in relation to the promoter was determined by restriction enzymes and sequencing. Then, this cassette was transferred to binary vector pZP211 and these cassettes were transferred into Agrobacterium tumefaciens LBA4404. S. lycopersicum cv. Micro-Tom (MT) and MT-Rg1 plants were transformed. In addition, seedlings were subjected to hormone treatments using a synthetic auxin (- naphthalene acetic acid) and cyclosporin A (cyclophilin inhibitor) treatments and it was found that the hormone treatment there were changes in development of lateral roots pattern, probably related to decreases in auxin signaling caused by reduction of LeCYP1 in MT-dgt plants while cyclosporin A treatments, there was a slight delay in flowering in cv. MT plants. Furthermore, assay with real-time PCR (RT-qPCR) were done for expression level analysis from LeCYP1 and ARP in order to functionally characterize these sequences in tomato plants.

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Sugarcane is an important culture for Brazil that holds almost half of all worldwide productivity. Plants face many challenges, because of biotic and abiotic stresses presents in the production field, which could prevent plants from reaching their genetic potential. As consequence, those stresses can generate Reactive Oxygen Species – ROS – that can cause damages on DNA. Another consequence of stress is the early-flowering process, which contributes for a reduction on yield. In this context, the aim of this work is to characterize ScMUTM1 and ScMUTM2, two DNA glycosylases belonging to base excision repair pathway; and identify genes potentially related to stress and DNA repair in two sugarcane cultivars with contrasting flowering phenotypes. The characterization of the DNA glycosylases included the construction of vector to over express the recombinant proteins ScMUTM1 and ScMUTM2; they will be used in a near future to purification of these proteins and use in enzymatic assays. It was also made a phylogenetic reconstruction of this gene in plants and analysis of its promoter. With the phylogenetic analysis, it is possible to observe the presence of these genes grouped inside a branch with monocots and another one with dicots. This suggests that the duplication of this gene probably occurred after the separation of these two groups. The analysis of the promotor of MUTM shows of the presence of stress-related regulatory motifs at ScMUTM2 promoter, when compared with ScMUTM1. This may suggests that ScMUTM1 might be suffering sub functionalization process. After the analysis of microarrays data, it is observed an up-regulation from some stress-related genes in one of the conditions analyzed, related to early flowering process.

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Sugarcane is an important culture for Brazil that holds almost half of all worldwide productivity. Plants face many challenges, because of biotic and abiotic stresses presents in the production field, which could prevent plants from reaching their genetic potential. As consequence, those stresses can generate Reactive Oxygen Species – ROS – that can cause damages on DNA. Another consequence of stress is the early-flowering process, which contributes for a reduction on yield. In this context, the aim of this work is to characterize ScMUTM1 and ScMUTM2, two DNA glycosylases belonging to base excision repair pathway; and identify genes potentially related to stress and DNA repair in two sugarcane cultivars with contrasting flowering phenotypes. The characterization of the DNA glycosylases included the construction of vector to over express the recombinant proteins ScMUTM1 and ScMUTM2; they will be used in a near future to purification of these proteins and use in enzymatic assays. It was also made a phylogenetic reconstruction of this gene in plants and analysis of its promoter. With the phylogenetic analysis, it is possible to observe the presence of these genes grouped inside a branch with monocots and another one with dicots. This suggests that the duplication of this gene probably occurred after the separation of these two groups. The analysis of the promotor of MUTM shows of the presence of stress-related regulatory motifs at ScMUTM2 promoter, when compared with ScMUTM1. This may suggests that ScMUTM1 might be suffering sub functionalization process. After the analysis of microarrays data, it is observed an up-regulation from some stress-related genes in one of the conditions analyzed, related to early flowering process.

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The flowering is a physiological process that it is vital for plants. This physiological process has been well studied in the plant model Arabidopsis, but in sugarcane this process is not well known. The transition of the shoot apical meristem from vegetative to flowering is a critical factor for plant development. At Brazil northeastern region, the transition to flowering in sugarcane has an important effect as it may reduce up to 60% its production. This is a consequence of the sugar translocation from stalks to the shoot apical meristem which is necessary during the flowering process. Therefore, the aim of this work was to explore and analyze cDNAs previously identified using subtractive cDNA libraries. The results showed that these cDNAs showed differential expression profile in varieties of sugarcane (early x late flowering). The in silico analysis suggested that these cDNAs had homology to calmodulin, NAC transcription factor and phosphatidylinositol, a SEC14, which were described in the literature as having a role in the process of floral development. To better understand the role of the cDNA homologous to calmodulin, tobacco plants were transformed with overexpression cassettes in sense and antissense orientation. Plants overexpressing the cassette in sense orientation did not flowered, while plants overexpressing the cassette in the antissense orientation produced flowers. The data obtained in this study suggested the possible role from CAM sequence, SEC14 and NAC in the induction/floral development pathway in sugarcane, this is the first study in order to analyze these genes in the sugarcane flowering process.

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Sugarcane is one of the most important products of the world and Brazil is responsible for 25 % of the world production. One problem of this culture at northeast of Brazil is the early flowering. In our laboratory, it has been made before four subtractive libraries using early and late flowering genotypes in order to identify messages related to the flowering process. In this work, two cDNAs were chosen to make in silico analysis and overexpression constructs. Another approach to understand the flowering process in sugarcane was to use proteomic tools. First, the protocol for protein extraction using apical meristem was set up. After that, these proteins were separated on two bidimensional gels. It was possible to observe some difference for some regions of these gels as well as some proteins that can be found in all conditions. The next step, spots will be isolated and sequence on MS spectrometry in order to understand this physiological process in sugarcane

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The IT capability is a organizational ability to perform activities of this role more effectively and an important mechanism in creating value. Its building process (stages of creation and development) occurs through management initiatives for improvement in the performance of their activities, using human resources and IT assets complementary responsible for the evolution of their organizational routines. This research deals with the IT capabilities related to SIG (integrated institutional management systems), built and deployed in UFRN (Universidade Federal do Rio Grande do Norte) to realization and control of administrative, academic and human resources activities. Since 2009, through cooperative agreements with federal and educational institutions of direct administration, UFRN has supported the implementation of these systems, currently involving more than 30 institutions. The present study aims to understand how IT capabilities, relevant in the design, implementation and dissemination of SIG, were built over time. This is a single case study of qualitative and longitudinal nature, performed by capturing, coding and analysis from secondary data and from semi-structured interviews conducted primarily with members of Superintenência de Informática, organizational unit responsible for SIG systems in UFRN. As a result, the technical, of internal relationship and external cooperation capabilities were identified as relevant in the successful trajectory of SIG systems, which have evolved in different ways. The technical capacity, initiated in 2004, toured the stages of creation and development until it reached the stage of stability in 2013, due to technological limits. Regarding the internal relationship capability, begun in 2006, it toured the stages of creation and development, having extended its scope of activities in 2009, being in development since then. Unlike the standard life cycle observed in the literature, the external cooperation capability was initiated by an intensity of initiatives and developments in the routines in 2009, which were decreasing to cease in 2013 in order to stabilize the technological infrastructure already created for cooperative institutions. It was still identified the start of cooperation in 2009 as an important event selection, responsible for changing or creating trajectories of evolution in all three capacities. The most frequent improvements initiatives were of organizational nature and the internal planning activity has been transformed over the routines of the three capabilities. Important resources and complementary assets have been identified as important for the realization of initiatives, such as human resources technical knowledge to the technical capabilities and external cooperation, and business knowledge, for all of them, as well as IT assets: the iproject application for control of development processes, and the document repository wiki. All these resources and complementary assets grew along the capacities, demonstrating its strategic value to SINFO/UFRN

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The growing importance of tourism in overall economic activity worldwide has favored the intensification of competition among cities that seek to create environments attractive to tourists and potential investors. It has been common practice to import characteristics of the business environment in the public management of cities. The city marketing is a key tool used by public leaders to promote a linkage between the tourism image and urban image and involves, in addition to promoting the image of the city, the planning of interventions in urban space, trying to formulate a positive image of the city able to facilitate the deployment of capital. This research seeks to understand the nature of city marketing as part of contemporary urban management and analyzes how is its application in decisions concerning the promotion of tourism in Natal/RN. The approach of this research is qualitative, exploratory and descriptive, in which respondents were the main leaders of two of the official tourism site, the Empresa Potiguar de Promoção Turística and the Secretaria Municipal de Turismo e Desenvolvimento Econômico. It was found that there is a strong articulation of public power with private enterprise in the design and conduct of the actions of urban marketing, that from the survey data show that the behavior of target markets provide guidelines for taking strategic decisions relating to tourism. Sun and sea are some key elements explored to form the image of Natal and to authorize the sale of the city as a tropical paradise. However, there is an increase in the diversification of tourism products, seeking to increase flow to the segments of ecotourism, adventure, business and culture. It s also growing the use of local culture as a tourism product, however, the cultural representation focuses on superficial values and does not bring to light the social and historical richness that the city has. Public authorities use the city marketing strategies as a means able to maximize the attractiveness of Natal urban space to investors, business groups and tourists. It can be observed that urban managers seek solutions that can continuously increase the tours, which often manifests in interventions that focus the tourist areas of the city, in oposition of those who do not contribute to a positive reading of the city, which ultimately generate the worsening of spacial and social inequalities

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Ce travaille de dissertation (Master) est le résultat de trois ans de travaille auprès de Mirandas, endroit situé à 22 kilomètres de la ville de Caraúbas, région ouest de l Etat du Rio Grande do Norte. Pendant cette période, on a compris que les habitants de Mirandas se identifient et sont identifiés pour les autres comme des caboclos , catégorie de classification que dans le discours courant, soit à Mirandas ou dans la siège de la municipalité, a des rapports avec la vision que les tous premiers habitants de Mirandas sont venus de la ville du Cabo , dans l Etat de Pernambuco. Et pourtant ; l usage qu a été fait de cette version a fini pour cacher des éléments qui ont défini l identité de Mirandas, tel comme la probable descendance indigène que parfois est actionnée dans les narratives qui montrent des rapports entre les portugais et des femmes indiens qu ont été pris à casco de cavalo (cheval) . (MACÊDO, 2010; CAVIGNAC, 2003). Donc, cette ethnographie porte à la lumière des éléments que sont utilisés pour les mirandenses pour définir leur identité. Cette identité peut être bien visualisé a partir des éléments que les caboclos font des signes diacritiques (BARTH, 1998; OLIVEIRA, 1976), bien comme des rapports établis entre les habitants de Mirandas et ces de Caraúbas. Des rapports qui ont rendu possible les manifestations d affinité et de conflits et que, donc, nous aide à entendre comme cette identité se réactualise dans ces rapports d union et opposition que les mirandenses construisent avec les caraubenses. Cela arrive dans les rapports territoriaux, économiques et de façon plus marquante, dans les rapports religieux.

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In view of a field research carried out by a team connected to the universe of body modification, it is possible to discern some uses and meanings linked to these forms of body interventions. Body modification or body change is part of the circuit of piercings and tattoos, although they are socially less thinned and more extreme, like scarifications, subcutaneous implants, bifurcated tongues, surfaces and body suspensions. The aim of this paper is to cast an anthropological glance on these practices, joining at the same context all the subjects involved with these techniques, placing them inside the same relational focus and capturing their journeys and trajectories. The discussions are concentrated on the notion of body building and urban life style. Ideas as personal distinctness and prestige imitation are also present in this universe, as well as matters attached to genre, pleasure, art, and to the so-called alternative circuit . This way, the ethnography so far presented here, reveals the complex and contemporaneous character of these practices of body markings in which the body appears as the central element in the experiences of the subjects of this study

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Le thème du patrimoine culturel architectural et urbain continue d avoir une place importante dans le milieu technique et scientifique. Le concept s est élargi et aujourd hui comprend différentes procédures de projets d intervention. L importance accordée au thème amène à l inclusion de la matière de techniques rétrospectives et aux contenus qui en sont liés: conservation, restauration, restructuration et reconstruction d édifices et ensembles urbains, dans les parcours des cours d architecture et d urbanisme au Brésil établies par le Ministère de l Education Nationale (MEC) dans les années quatre-vingt-dix, postérieurement incorporés dans les directrices disciplinaires nationales. Nous partons des discussions théoriques et conceptuelles du Domaine du Patrimoine Culturel, ainsi que des principales théories pédagogiques d enseignements et d apprentissage articulées au projet. Dans ce contexte les objectifs principaux de cette thèse consistent à systématiser et à analyser les principales procédures méthodologiques contribuant pour la construction de méthodes d enseignement tournée vers des activités pratiques dans ce domaine. Pour cela, la recherche a été systématisée dans une approche à deux niveaux. En ce qui concerne le premier, basé sur des données secondaires, neuf cours d architecture et urbanisme ont étés identifiés entre institutions publiques d enseignement supérieur dont huit brésiliennes et une française, considérées représentatives en ce qui concerne les pratiques d enseignement de projet et de patrimoine culturel. Trente disciplines dédiées à la matière ont été également reconnues initialement, et postérieurement, cinq disciplines qui possèdent un emploi du temps dédié à la pratique de projet ont aussi été reconnues. Dans le deuxième cas, basée sur des données primaires, ont étés analysées les méthodologies et les stratégies d enseignement de projet basées sur les définitions des matières et des autres éléments des plans de travail avec des observations, des entrevues et des questionnaires en trois ateliers. Par rapport aux résultats nous avons constaté que toutes les écoles possèdent les contenus de la matière, mais peu d entre elles privilégient la relation du projet appliqué au patrimoine culturel. Nous avons constaté que les questions des projets dans ce contexte, même s elles sont considérées complexes, ont privilégié le listage et l analyse du site. L atelier qui intègre les fondements des théories de préservation, l histoire de l architecture et urbanisme et techniques anciennes et actuelles, est mis en valeur comme un modèle cohérent avec les propositions d intégration des connaissances théoriques et pratiques du projet appliqué à la discipline. Basé sur ces constatations il est possible de démontrer quatre étapes du projet appliqué au patrimoine culturel: 1ª) les fondements généraux qui concernent les bases théoriques sur la préservation, histoire et technique rétrospective, par exemple, l appropriation de lois et normes et la sensibilisation de l élève sur les questions de patrimoine culturel; 2ª) le contacte avec la réalité qui inclut l appropriation du problème à partir de ces acteurs, de ces échelles, de cette lecture de site et l analyse de l objet d étude; 3ª) le développement de la proposition qui inclut programmes (fonctions existantes et propositions), définitions du partit (types d intervention), conception (hypothèse et discussion) et définition de proposition; 4ª) la finalisation du projet qui consiste à développer la proposition avec sa représentation graphique et sa présentation finale. Nous concluons que le projet en Domaine du Patrimoine Culturel demande une attention spéciale et doit être présent dans les cursus considérant les principes généraux nécessaires à la formation de l élève. Le binôme projet / patrimoine signifie avoir dans le cursus universitaire les contenus et questions nécessaires les connaissances, les variables et possibilités existantes dans le projet appliqué au patrimoine culturel de façon à ce que ces connaissances soient incorporées dans l exercice de projet et n apparaissent pas comme un simple contenu théorique sans articulation avec la pratique. Naturellement ces conclusions n épuisent pas la réflexion sur la question. Nous espérons que les analyses faites contribuent à définir des méthodologies d enseignements capables d êtres vérifiées et testées dans la pratique en salle de cours, et puisse collaborer avec les nouvelles recherches surtout celles qui ont pour but des nouvelles théories pédagogiques d enseignement apprentissage du projet en Domaine du Patrimoine Culturel

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The Sectoral Plan was inserted as an urban instrument in Master Plan of Natal (LC82/07), since then it has been deepened. It is guided to the equitable distribution of the benefits of urbanization, aimed the efetivation of the Right to the City and the Environmental function of property in the context of urban planning. Thereby, the Sectoral Plan was inserted into the perspective of promoting a favorable field for applying articulated of instruments defined in the Master Plan, and them with urban interventions, through the deepening of macrozoning - coming out of dimension of the city and going to a small fraction of the urban by reducing scaling - facilitating democratic management in accordance with the City Statute. However, the absence of regulation and the conceptual and operational fragility of the instrument, approached it of other existent experiences and instruments, limiting its application and evaluation. Considering the innovative nature of the instrument and the intense social participation, we inquire about conceptual and operational elements which could give greater effectiveness to Sector Plan in construction? Guided by that question, this dissertation aim to understand the nature and operational procedures of Sectoral Plan, seeking to do indications about the conceptual and operational aspects of implementing a Sector Plan. The neighborhood of Ponta Negra was selected as empirical universe because it was the first sector defined in the Master Plan. This research adopts reference authors who´s make reflection and insights about the matrix of Urban Reform, whose principles constitute the basis of the trajectory of revision of urban planning in Brazil since the 1980s, highlighting Raquel Rolnik, Nelson Saule Júnior and Orlando Alves dos Santos Júnior. For the local monitoring, we use the authors Alexsandro Ferreira Cardoso Silva, Rosa Maria Pinheiro Oliveira e Marise Costa da Souza Duarte, in order to understand the growth dynamics of Natal, lawfully and urbanistically

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At a time of changes on the territory during the 19th century, the political and socioeconomic elites of the province and later State of Rio Grande do Norte evolved a discourse in order to justify the permanence of Natal as a city holding a status of capital. In this work we analyze the means employed by the ruling classes to impose their wish to raise Natal to an outstanding position among the existing cities by intervening on the territory during a period of one hundred years (1820-1920). During that time, which was characterized by changing commercial flows and technological development, the elites interventions were essentially directed to the implementation of modes of transportation, especially the railway. We try to understand the reinforcement of Natal as a capital city not only in political and administrative terms, but mainly in a commercial and symbolic manner, through the discourse and interventions undertaken by the local administrative elites, who stimulated the creation of a set of relations on the territory that also imprinted visible marks in the capital s urban fabric. These interventions were based upon the establishment of an infrastructure for exporting the State s production, firstly through and despite the Potengi River, and later on by the construction of railways. Although the project of Natal s hegemony had been outlined before the establishment of the railway network, in both cases the ultimate objective was to reinforce and develop the capital city as a commercial urban center to the detriment of other cities

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The so-called "residential tourism" came to intensify the previous occupation of the coastal zones, characterized by traditional beach houses, and brought significant consequences for their spatial configuration, and especially for its scenic value of the landscape. Although there is the intention to regulate the activities of enterprising groups by some legal instruments to control the use and occupation of land, and to contain some negative effects, the actions of government are still inefficient in trying to follow the implications on the landscape from the accelerated growth of the real estate and touristic sectors. Supported in the speech of economic development and income generation, public managers prioritize areas to attract tourists to the detriment of preserving important physical attributes of the natural environment that contribute significantly to the quality of life. The result can be noticed in the use of natural elements as one of the major components in the land valorization, and in the immediate attraction of investors and enterprising. Therefore, the objective of this work is to contribute to the debate on the landscape preservation a little detailed thematic in view of their relevance in the current context - by indicating subsidies to the creation of a methodology for the evaluation and protection of coastal zones that may assist the government in creating new instruments, and better prepare it in control of the occupation of these areas. For such, was followed two ways to analysis: the indirect method and the direct method. The first is grounded in the evaluation of landscape attributes, which is based on the work of Raquel Tardin (2008) and Eduardo Cuesta, Encarnación Algarra and Isabel Pastor (2001). The second, based on the research of Leticia and Carlos Hardt (2010) and on the concepts of phenomenology - expressed by Antonio Christofoletti (1985) and Yi-Fu Tuan (1983) - considers the population perspective on the quality of the natural scenery. Developed through cartographic materials, photographic collections and quantitative tables, this dissertation utilized as a case study the beaches of Barra de Tabatinga and Camurupim, located in the city of Nísia Floresta/RN. Despite already being sighted spaces of advanced stage of landscape degradation in these locations, areas of remarkable scenic value can still be found, what reinforce the urgency in adopting preservationists actions. The absence of laws focused on the management and protection of the landscape singularities associated with the inefficiency of the government to invigilate the land occupation in coastal zones, encourage the excessive action of the real estate-tourism, and consequently make the government the main responsible for the environmental and landscape impacts in these areas - by its omission or by their connivance. Therefore, the legislation permeates this entire process and constitutes itself as the most effective way to guarantee the right to the landscape to present and future generations. Are also pointed some important considerations to build a methodology, especially concerning possibilities of improvements and adaptations of its applicability in each case

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The present dissertation has begun with an investigation about the application of the Technique of the Corporeal Mimesis, systematized by the Lume, Interdisciplinary Center for Theatrical Research of Unicamp, associated with the matrix of Rural Maracatu, specifically of the group Cambinda Brasileira (Nazaré da Mata PE), with the purpose of promote the training and the composition work of the actor. Starting from an empirical methodology, which used the analysis of practical experiences the application process of the Corporeal Mimesis and the construction of a theater play associated with bibliographical research, I could investigate possible ways of re-signification of this technique, using components related to the ritual and the game present in the popular event in question, and experience such paths through the conception of the play Cravo do Canavial